WO2011162968A1 - Agents médicinaux liés à un dipeptide - Google Patents

Agents médicinaux liés à un dipeptide Download PDF

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Publication number
WO2011162968A1
WO2011162968A1 PCT/US2011/039755 US2011039755W WO2011162968A1 WO 2011162968 A1 WO2011162968 A1 WO 2011162968A1 US 2011039755 W US2011039755 W US 2011039755W WO 2011162968 A1 WO2011162968 A1 WO 2011162968A1
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Prior art keywords
alkyl
group
prodrug
methyl
dipeptide
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PCT/US2011/039755
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English (en)
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Richard D. Dimarchi
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Indiana University Research And Technology Corporation
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Application filed by Indiana University Research And Technology Corporation filed Critical Indiana University Research And Technology Corporation
Priority to US13/697,050 priority Critical patent/US20130137849A1/en
Priority to CA2796879A priority patent/CA2796879A1/fr
Priority to JP2013516595A priority patent/JP6101202B2/ja
Priority to RU2013102991/04A priority patent/RU2013102991A/ru
Priority to MX2012014573A priority patent/MX2012014573A/es
Priority to CN201180031396.0A priority patent/CN102958533B/zh
Priority to KR1020127033201A priority patent/KR20130083843A/ko
Priority to EP11798607.5A priority patent/EP2588127A4/fr
Publication of WO2011162968A1 publication Critical patent/WO2011162968A1/fr
Priority to US15/016,959 priority patent/US20160158375A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22—Hormones
    • A61K38/28—Insulins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/08—Peptides having 5 to 11 amino acids
    • A61K38/095—Oxytocins; Vasopressins; Related peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
    • A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
    • A61K47/65—Peptidic linkers, binders or spacers, e.g. peptidic enzyme-labile linkers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P5/00—Drugs for disorders of the endocrine system
    • A61P5/14—Drugs for disorders of the endocrine system of the thyroid hormones, e.g. T3, T4
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/575—Hormones
    • C07K14/62—Insulins
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2/00—Peptides of undefined number of amino acids; Derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06008—Dipeptides with the first amino acid being neutral
    • C07K5/06017—Dipeptides with the first amino acid being neutral and aliphatic
    • C07K5/06026—Dipeptides with the first amino acid being neutral and aliphatic the side chain containing 0 or 1 carbon atom, i.e. Gly or Ala
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06008—Dipeptides with the first amino acid being neutral
    • C07K5/06017—Dipeptides with the first amino acid being neutral and aliphatic
    • C07K5/06034—Dipeptides with the first amino acid being neutral and aliphatic the side chain containing 2 to 4 carbon atoms
    • C07K5/06043—Leu-amino acid
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06008—Dipeptides with the first amino acid being neutral
    • C07K5/06017—Dipeptides with the first amino acid being neutral and aliphatic
    • C07K5/06034—Dipeptides with the first amino acid being neutral and aliphatic the side chain containing 2 to 4 carbon atoms
    • C07K5/06052—Val-amino acid
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06008—Dipeptides with the first amino acid being neutral
    • C07K5/06017—Dipeptides with the first amino acid being neutral and aliphatic
    • C07K5/0606—Dipeptides with the first amino acid being neutral and aliphatic the side chain containing heteroatoms not provided for by C07K5/06086 - C07K5/06139, e.g. Ser, Met, Cys, Thr
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06008—Dipeptides with the first amino acid being neutral
    • C07K5/06017—Dipeptides with the first amino acid being neutral and aliphatic
    • C07K5/0606—Dipeptides with the first amino acid being neutral and aliphatic the side chain containing heteroatoms not provided for by C07K5/06086 - C07K5/06139, e.g. Ser, Met, Cys, Thr
    • C07K5/06069—Ser-amino acid
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06086—Dipeptides with the first amino acid being basic
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06086—Dipeptides with the first amino acid being basic
    • C07K5/06095—Arg-amino acid
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06104—Dipeptides with the first amino acid being acidic
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06104—Dipeptides with the first amino acid being acidic
    • C07K5/06113—Asp- or Asn-amino acid
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06139—Dipeptides with the first amino acid being heterocyclic
    • C07K5/06147—Dipeptides with the first amino acid being heterocyclic and His-amino acid; Derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06139—Dipeptides with the first amino acid being heterocyclic
    • C07K5/06156—Dipeptides with the first amino acid being heterocyclic and Trp-amino acid; Derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06139—Dipeptides with the first amino acid being heterocyclic
    • C07K5/06165—Dipeptides with the first amino acid being heterocyclic and Pro-amino acid; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • peptide-based drugs can be used as highly effective medicines, they typically have relatively short duration of action and variable therapeutic index.
  • Mechanisms for providing extended release and an enhanced therapeutic index include sequestering molecules at the injection site or the use of prodrug derivative forms of the pharmaceutical, wherein the prodrug derivative is designed to delay onset of action and extend the half life of the drug.
  • the delayed onset of action is advantageous in that it allows systemic distribution of the prodrug prior to its activation. Accordingly, the administration of prodrugs eliminates complications caused by peak activities upon administration and increases the therapeutic index of the parent drug.
  • Receptor recognition and subsequent processing of peptide and protein agonists is the primary route of degradation of many peptide and protein-based drugs.
  • binding of the peptide drug to its receptor will result in biological stimulation, but will also initiate the subsequent deactivation of the peptide/protein induced pharmacology through the enzymatic degradation of the peptide or protein.
  • existing pharmaceutical compounds can be modified to prevent their interaction with their corresponding receptor.
  • drugs can be modified by the linkage of a non- enzymatic self cleaving dipeptide to the drug to form a complex that functions either as a depot composition, to localize the drug at the injection site for release in a controlled manner, or as a prodrug that is distributed through out the body but incapable of interacting with its receptor.
  • a non-enzymatic self cleaving dipeptide moiety that can be covalently linked to a medicinal agent, wherein the dipeptide (and any compound linked to the dipeptide) is released from the medicinal agent at a predetermined length of time after exposure to physiological conditions.
  • the rate of cleavage depends on the structure and stereochemistry of the dipeptide element and also on the strength of the nucleophile present on the dipeptide that induces cleavage and diketopiperazine or diketomorpholine formation.
  • a complex comprising a known drug and a dipeptide of the structure A-B is provided, wherein A is an amino acid or a hydroxyl acid and B is an N-alkylated amino acid that is linked to the drug through formation of an amide bond between B and an amine of the drug.
  • the amino acids of the dipeptide are selected such that a non-enzymatic chemical cleavage of A-B from the drug produces a diketopiperazine or diketomorpholine and the reconstituted native drug.
  • A is an amino acid or a hydroxyl acid
  • B is an N-alkylated amino acid
  • Q is a an amine bearing medicinal agent
  • the dipeptide A-B further comprises a depot polymer linked to the side chain of A or B, and said dipeptide is linked to Q through formation of an amide bond between A-B and an amine of Q.
  • the depot polymer is selected to be of a sufficient size that the complex A-B-Q is effectively sequestered at the site of injection or is otherwise incapable of interacting with its target (e.g., receptor).
  • Chemical cleavage of A-B from Q produces a diketopiperazine or diketomorpholine and releases the active drug to the patient in a controlled manner over a predetermined duration of time after administration.
  • prodrug derivatives of known pharmaceutical agents are prepared to extend the peptide or protein's biological half life based on a strategy of inhibiting recognition of the prodrug by the corresponding receptor.
  • the prodrugs disclosed herein will ultimately be chemically converted to structures that can be recognized by the receptor, wherein the speed of this chemical conversion will determine the time of onset and duration of in vivo biological action.
  • the molecular design disclosed in this application relies upon an intramolecular chemical reaction that is not dependent upon additional chemical additives, or enzymes.
  • the prodrug derivative is prepared by covalently linking a dipeptide element to an active site of the medicinal agent via an amide linkage.
  • the dipeptide is covalently bound to the medicinal agent at a position that interferes with the medicinal agent's ability to interact with its corresponding receptor or cofactor.
  • the dipeptide element is linked to the N-terminus of a bioactive peptide. Subsequent removal of the dipeptide, under physiological conditions and in the absence of enzymatic activity, restores full activity to the polypeptide.
  • a prodrug having the general structure of A-B- Q.
  • Q is a medicinal agent, including for example a bioactive peptide.
  • Q is selected from the group of nuclear hormones consisting of thyroid hormone, estrogen, testosterone, and glucocorticoid, as well as analogs, derivatives and conjugates of the foregoing, and A-B represents a dipeptide prodrug linked to Q through an amide bond. More particularly, in one embodiment A is an amino acid or a hydroxyl acid and B is an N-alkylated amino acid linked to Q through formation of an amide bond between A-B and an amine of Q.
  • the chemical cleavage half-life (t ⁇ ) of A-B from Q is at least about 1 hour to about 1 week in PBS under physiological conditions.
  • Q comprises an amino acid sequence
  • A, B, or the amino acid of Q to which A-B is linked is a non-coded amino acid
  • chemical cleavage of A-B from Q is at least about 90% complete within about 1 to about 720 hours in PBS under physiological conditions.
  • a and B are selected to inhibit enzymatic cleavage of the A-B dipeptide from Q by enzymes found in mammalian serum.
  • a and/or B are selected such that the cleavage half-life of A-B from Q in PBS under physiological conditions is not more than two fold the cleavage half-life of A-B from Q in a solution comprising a DPP-IV protease (i.e., cleavage of A-B from Q does not occur at a rate more than 2x faster in the presence of DPP-IV protease and
  • a and/or B is an amino acid in the D stereoisomer configuration.
  • A is an amino acid in the D stereoisomer configuration and B is an amino acid in the L stereoisomer configuration.
  • A is an amino acid in the L stereoisomer configuration and B is an amino acid in the D stereoisomer configuration.
  • A is an amino acid in the D stereoisomer configuration and B is an amino acid in the D stereoisomer configuration.
  • A is an amino acid in the D stereoisomer configuration and B is an amino acid in the D stereoisomer configuration.
  • R 2> P4 and R 8 are independently selected from the group consisting of H, Ci-Cie alkyl, C 2 -C 18 alkenyl, (Ci-Cie alkyl)OH, (Ci-Cie alkyl)SH, (C 2 -C 3 alkyl)SCH 3 , (Ci-C 4 alkyl)CONH 2 , (C1-C4 alkyl)COOH, (C1-C4 alkyl)NH 2 , (C1-C4
  • R 5 is NHR 6 or OH
  • R 6 is H, CrC 8 alkyl or R 6 and R 2 together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;
  • R 7 is selected from the group consisting of H and OH.
  • R ⁇ R 2> R ⁇ and R 8 are independently selected from the group consisting of H, C1-C18 alkyl, C 2 -C 18 alkenyl, (CrC 18 alkyl)OH, (CrC 18 alkyl)SH, (C 2 -C 3 alkyl)SCH 3 , (C1-C4 alkyl)CONH 2 , (C1-C4 alkyl)COOH, (C1-C4 alkyl)NH 2 , (C1-C4
  • R 3 is selected from the group consisting of CrC 18 alkyl, (CrC 18 alkyl)OH, (Ci-Cie alkyl)NH 2 , (CrC 18 alkyl)SH, (C 0 -C 4 alkyl)(C 3 -C 6 )cycloalkyl, (C 0 -C 4 alkyl)(C 2 -C 5 heterocyclic), (C 0 -C 4 alkyl)(C 6 -C 10 aryl)R 7 , and (C1-C4 alkyl)(C 3 -C 9 heteroaryl) or R 4 and R together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;
  • R 5 is NHR 6 or OH
  • Re is H, CrC 8 alkyl or R 6 and R together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring; and R 7 is selected from the group consisting of hydrogen, Q-Cig alkyl, C 2 -Ci 8 alkenyl, (C 0 -C 4 alkyl)CONH 2> (C 0 -C 4 alkyl)COOH, (C 0 -C 4 alkyl)NH 2 , (C 0 -C 4 alkyl)OH, and halo.
  • amino acid encompasses any molecule containing both amino and carboxyl functional groups, wherein the amino and carboxylate groups are attached to the same carbon (the alpha carbon).
  • the alpha carbon optionally may have one or two further organic substituents.
  • An amino acid can be designated by its three letter code, one letter code, or in some cases by the name of its side chain.
  • an unnatural amino acid comprising a cyclohexane group attached to the alpha carbon is termed "cyclohexane” or "cyclohexyl.”
  • designation of an amino acid without specifying its stereochemistry is intended to encompass either the L or D form of the amino acid, or a racemic mixture.
  • an amino acid is designated by its three letter code and includes a superscript number (i.e., Lys -1 )
  • a designation is intended to specify the native L form of the amino acid
  • the D form will be specified by inclusion of a lower case d before the three letter code and superscript number (i.e., dLys "1 ).
  • hydroxyl acid refers to amino acids that have been modified to replace the alpha carbon amino group with a hydroxyl group.
  • non-coded amino acid encompasses any amino acid that is not an L-isomer of any of the following 20 amino acids: Ala, Cys, Asp, Glu, Phe, Gly, His, He, Lys, Leu, Met, Asn, Pro, Gin, Arg, Ser, Thr, Val, Trp, Tyr.
  • a “dipeptide” is the result of the linkage of an alpha amino acid or an alpha hydroxyl acid to another amino acid, through a peptide bond.
  • chemical cleavage absent any further designation encompasses a non-enzymatic reaction that results in the breakage of a covalent chemical bond.
  • a “bioactive peptide” refers to peptides which are capable of exerting a biological effect in vitro and/or in vivo.
  • a general reference to a peptide is intended to encompass peptides that have modified amino and carboxy termini.
  • an amino acid sequence designating the standard amino acids is intended to encompass standard amino acids at the N- and C- terminus as well as a corresponding hydroxyl acid at the N-terminus and/or a corresponding C-terminal amino acid modified to comprise an amide group in place of the terminal carboxylic acid.
  • an "acylated" amino acid is an amino acid comprising an acyl group which is non-native to a naturally- occurring amino acid, regardless by the means by which it is produced.
  • exemplary methods of producing acylated amino acids and acylated peptides are known in the art and include acylating an amino acid before inclusion in the peptide or peptide synthesis followed by chemical acylation of the peptide.
  • the acyl group causes the peptide to have one or more of (i) a prolonged half-life in circulation, (ii) a delayed onset of action, (iii) an extended duration of action, (iv) an improved resistance to proteases, such as DPP-IV, and (v) increased potency at a medicinal agent peptide receptor.
  • an "alkylated” amino acid is an amino acid comprising an alkyl group which is non-native to a naturally- occurring amino acid, regardless of the means by which it is produced.
  • Exemplary methods of producing alkylated amino acids and alkylated peptides are known in the art and including alkylating an amino acid before inclusion in the peptide or peptide synthesis followed by chemical alkylation of the peptide.
  • alkylation of peptides will achieve similar, if not the same, effects as acylation of the peptides, e.g., a prolonged half-life in circulation, a delayed onset of action, an extended duration of action, an improved resistance to proteases, such as DPP-IV, and increased potency at a medicinal agent peptide receptors.
  • prodrug is defined as any compound that undergoes chemical modification before exhibiting its pharmacological effects.
  • immediate agents refers to a biologically active substance or substances that mediate their effect through interacting with a receptor
  • medicinal agents are defined as compounds falling into one of four classes:
  • non-glucagon and non-insulin peptide-based hormones and derivatives 3. proteins within the class of 4-helix bundle proteins, including for example growth hormone, leptin, erythropoietin, colony stimulating factors (such as GCSF) and interferons; and.
  • blood clotting factors including for example, tissue plasminogen activators (TP A), Factor VII, Factor VIII and Factor IX.
  • nuclear hormone is a compound that when bound to its corresponding receptor, will directly interact with and control the expression of genomic DNA.
  • nuclear hormones include thyroid hormone,
  • glucocorticoids glucocorticoids, estrogens, androgens, vitamin A and vitamin D.
  • a "receptor” is a molecule that recognizes and binds with specific molecules in a high affinity interaction, producing some effect (either directly or indirectly) in a cell, or on the cells and/or tissues of the host organism.
  • a "cellular receptor” is a molecule on or within a cell that recognizes and binds with specific molecules, producing some effect (either directly or indirectly) in the cell.
  • non-glucagon and non-insulin peptide-based hormone is a hormone that comprises a peptide sequence, but specifically excludes insulin, insulin derivatives and analogs that specifically bind to the insulin receptor, insulin-like growth factors (IGFs) and glucagon superfamily peptides.
  • IGFs insulin-like growth factors
  • identity as used herein relates to the similarity between two or more sequences. Identity is measured by dividing the number of identical residues by the total number of residues and multiplying the product by 100 to achieve a percentage. Thus, two copies of exactly the same sequence have 100% identity, whereas two sequences that have amino acid deletions, additions, or substitutions relative to one another have a lower degree of identity.
  • BLAST Basic Local Alignment Search Tool, Altschul et al. (1993) J. Mol. Biol. 215:403-410) are available for determining sequence identity.
  • glucagon related peptide is directed to those peptides which have biological activity (as agonists or antagonists) at any one or more of the glucagon, GLP-1, GLP-2, and GIP receptors and comprise an amino acid sequence that shares at least 40% sequence identity (e.g., 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%) with at least one of native glucagon (SEQ ID NO: 1), native
  • oxyntomodulin SEQ ID NO: 51
  • native exendin-4 SEQ ID NO: 54
  • native GLP-1 SEQ ID NO: 50
  • native GLP-2 SEQ ID NO: 53
  • native GIP SEQ ID NO: 52
  • glucagon superfamily refers to a group of peptides related in structure in their N-terminal and C-terminal regions (see, for example, Sherwood et al., Endocrine Reviews 21: 619-670 (2000)). Members of this group include all glucagon related peptides, as well as Growth Hormone Releasing Hormone (GHRH; SEQ ID NO: 8), vasoactive intestinal peptide (VIP; SEQ ID NO: 55), Pituitary adenylate cyclase-activating polypeptide 27 (PACAP-27; SEQ ID NO: 56), peptide histidine isoleucine (PHI), peptide histidine methionine (PHM; SEQ ID NO: 57), and Secretin (SEQ ID NO: 58), and analogs, derivatives or conjugates with up to 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acid modifications relative to the native peptide.
  • GHRH Growth Hormone Releasing Hormone
  • the term "pharmaceutically acceptable carrier” includes any of the standard pharmaceutical carriers, such as a phosphate buffered saline solution, water, emulsions such as an oil/water or water/oil emulsion, and various types of wetting agents.
  • the term also encompasses any of the agents approved by a regulatory agency of the US Federal government or listed in the US Pharmacopeia for use in animals, including humans.
  • PBS phosphate buffered saline
  • standard PBS refers to a solution having have a final concentration of 137 mM NaCl, 10 mM Phosphate, 2.7 mM KCl, and a pH of 7.2-7.4.
  • pharmaceutically acceptable salt refers to salts of compounds that retain the biological activity of the parent compound, and which are not biologically or otherwise undesirable. Many of the compounds disclosed herein are capable of forming acid and/or base salts by virtue of the presence of amino and/or carboxyl groups or groups similar thereto.
  • Pharmaceutically acceptable base addition salts can be prepared from inorganic and organic bases.
  • Salts derived from inorganic bases include by way of example only, sodium, potassium, lithium, ammonium, calcium and magnesium salts.
  • Salts derived from organic bases include, but are not limited to, salts of primary, secondary and tertiary amines.
  • Pharmaceutically acceptable acid addition salts may be prepared from inorganic and organic acids. Salts derived from inorganic acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like.
  • Salts derived from organic acids include acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluene-sulfonic acid, salicylic acid, and the like.
  • treating includes prophylaxis of the specific disorder or condition, or alleviation of the symptoms associated with a specific disorder or condition and/or preventing or eliminating said symptoms.
  • an "effective” amount or a “therapeutically effective amount” of a drug refers to a nontoxic but sufficient amount of the drug to provide the desired effect.
  • the amount that is “effective” will vary from subject to subject, depending on the age and general condition of the individual, mode of administration, and the like. Thus, it is not always possible to specify an exact “effective amount.” However, an appropriate “effective” amount in any individual case may be determined by one of ordinary skill in the art using routine experimentation.
  • parenteral means not through the alimentary canal but by some other route such as subcutaneous, intramuscular, intraspinal, or intravenous.
  • an amino acid “modification” refers to a substitution, addition or deletion of an amino acid, and includes substitution with, or addition of, any of the 20 amino acids commonly found in human proteins, as well as atypical or non- naturally occurring amino acids.
  • Commercial sources of atypical amino acids include Sigma- Aldrich (Milwaukee, WI), ChemPep Inc. (Miami, FL), and Genzyme
  • Atypical amino acids may be purchased from commercial suppliers, synthesized de novo, or chemically modified or derivatized from naturally occurring amino acids.
  • Amino acid modifications include linkage of an amino acid to a conjugate moiety, such as a hydrophilic polymer, acylation, alkylation, and/or other chemical derivatization of an amino acid.
  • substitution refers to the replacement of one amino acid residue by a different amino acid residue.
  • conservative amino acid substitution is defined herein as exchanges within one of the following five groups: I. Small aliphatic, nonpolar or slightly polar residues:
  • polyethylene glycol chain refers to mixtures of condensation polymers of ethylene oxide and water, in a branched or straight chain, represented by the general formula H(OCH 2 CH 2 ) k OH, wherein k is at least 9. Absent any further characterization, the term is intended to include polymers of ethylene glycol with an average total molecular weight selected from the range of 500 to 60,000 Daltons. "Polyethylene glycol chain” or “PEG chain” is used in combination with a numeric suffix to indicate the approximate average molecular weight thereof. For example, PEG-5,000 (5k PEG ) refers to polyethylene glycol chain having a total molecular weight average of about 5,000 Daltons.
  • pegylated refers to a compound that has been modified from its native state by linking a polyethylene glycol chain to the compound.
  • a “pegylated polypeptide” is a polypeptide that has a PEG chain covalently bound to the polypeptide.
  • Linker is a bond, molecule or group of molecules that binds two separate entities to one another. Linkers may provide for optimal spacing of the two entities or may further supply a labile linkage that allows the two entities to be separated from each other. Labile linkages include photocleavable groups, acid-labile moieties, base-labile moieties and enzyme-cleavable groups.
  • a "dimer” is a complex comprising two subunits covalently bound to one another via a linker.
  • dimer when used absent any qualifying language, encompasses both homodimers and heterodimers.
  • a homodimer comprises two identical subunits, whereas a heterodimer comprises two subunits that differ, although the two subunits are substantially similar to one another.
  • C -C n alkyl wherein n can be from 1 through 6, as used herein, represents a branched or linear alkyl group having from one to the specified number of carbon atoms.
  • Typical CrC 6 alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, iso-propyl, butyl, iso-butyl, sec-butyl, tert-butyl, pentyl, hexyl and the like.
  • C2-C n alkenyl wherein n can be from 2 through 6, as used herein, represents an olefinically unsaturated branched or linear group having from 2 to the specified number of carbon atoms and at least one double bond.
  • C 2 -C n alkynyl wherein n can be from 2 to 6, refers to an unsaturated branched or linear group having from 2 to n carbon atoms and at least one triple bond. Examples of such groups include, but are not limited to, 1-propynyl, 2- propynyl, 1-butynyl, 2-butynyl, 1-pentynyl, and the like.
  • aryl refers to a mono- or bicyclic carbocyclic ring system having one or two aromatic rings including, but not limited to, phenyl, naphthyl, tetrahydronaphthyl, indanyl, indenyl, and the like.
  • the size of the aryl ring and the presence of substituents or linking groups are indicated by designating the number of carbons present.
  • (Ci-C 3 alkyl) (C6-C 10 aryl) refers to a 6 to 10 membered aryl that is attached to a parent moiety via a one to three membered alkyl chain.
  • heteroaryl refers to a mono- or bi- cyclic ring system containing one or two aromatic rings and containing at least one nitrogen, oxygen, or sulfur atom in an aromatic ring.
  • the size of the heteroaryl ring and the presence of substituents or linking groups are indicated by designating the number of carbons present.
  • (C C n alkylXCs-Ce heteroaryl) refers to a 5 or 6 membered heteroaryl that is attached to a parent moiety via a one to "n" membered alkyl chain.
  • halo refers to one or more members of the group consisting of fluorine, chlorine, bromine, and iodine.
  • charged amino acid refers to an amino acid that comprises a side chain that is negatively charged (i.e., de-protonated) or positively charged (i.e., protonated) in aqueous solution at physiological pH.
  • negatively charged amino acids include aspartic acid, glutamic acid, cysteic acid, homocysteic acid, and homoglutamic acid
  • positively charged amino acids include arginine, lysine and histidine.
  • Charged amino acids include the charged amino acids among the 20 amino acids commonly found in human proteins, as well as atypical or non-naturally occurring amino acids.
  • acidic amino acid refers to an amino acid that comprises a second acidic moiety (i.e. other than the cc-carboyxl group that all amino acids possess), including for example, a carboxylic acid or sulfonic acid group.
  • patient without further designation is intended to encompass any warm blooded vertebrate domesticated animal (including for example, but not limited to livestock, horses, cats, dogs and other pets) and humans.
  • a method for increasing an administered drug's duration of action and improving its therapeutic index.
  • the method comprises linking a dipeptide element to the drug via an amide linkage to produce a dipeptide/drug complex that is either sequestered at its point of
  • two or more dipeptide elements are linked via an amide bond to the drug.
  • the dipeptide will be cleaved via a non-enzymatic degradation mechanism thus releasing the active drug for interaction with its target.
  • the rate of cleavage depends on the structure and stereochemistry of the dipeptide element and also on the strength of the nucleophile present on the dipeptide that induces cleavage and diketopiperazine or diketomorpholine formation.
  • the non- enzymatic half time (tl/2) of the dipeptide/drug complex can be selected to be between 1-720 hrs under physiological conditions.
  • Physiological conditions as disclosed herein are intended to include a temperature of about 35 to 40 °C and a pH of about 7.0 to about 7.4, and more typically include a pH of 7.2 to 7.4 and a temperature of 36 to 38 °C. Since physiological pH and temperature are tightly regulated within a highly defined range, the speed of conversion from dipeptide/drug complex to drug will exhibit high intra and interpatient reproducibility.
  • the dipeptide element is covalently bound to the drug via an amide linkage at an active site of the drug to form a prodrug derivative of the drug.
  • the prodrug will exhibit no more than 10% of the activity of the parent drug, in one embodiment the prodrug exhibits less than 10%, less than 5%, about 1%, or less than 1% activity relative to the parent drug.
  • the prodrugs disclosed herein will ultimately be chemically converted to structures that can be recognized by the native receptor of the drug, wherein the speed of this chemical conversion will determine the time of onset and duration of in vivo biological action.
  • the drug is a medicinal agent.
  • the molecular design disclosed in this application relies upon an intramolecular chemical reaction that is not dependent upon additional chemical additives, or enzymes, wherein the speed of conversion is controlled by the chemical nature of the dipeptide substituents.
  • the dipeptide element is covalently bound to the drug via an amide linkage, and the dipeptide further comprises a depot polymer linked to dipeptide.
  • the drug is a medicinal agent.
  • two or more depot polymers are linked to a single dipeptide element.
  • the depot polymer is selected to be biocompatible and of sufficient size that the drug modified by covalent attachment of the dipeptide remains sequestered at an injection site and/or incapable of interacting with its corresponding receptor upon administration to a patient. Subsequent cleavage of the dipeptide releases the drug to interact with its intended target. Selection of different combinations of substituents on the dipeptide element will allow for the preparation of injectable compositions that comprise a mixture of dipeptide/drug complexes that release the drug over a desired time frame.
  • any known pharmaceutical that comprises a primary or secondary amine, or that can be modified to comprise such an amine without loss of function can be modified to comprise a dipeptide element that will cleave via an intramolecular chemical reaction that is not dependent upon additional chemical additives, or enzymes.
  • a cleavage will regenerate the structure of the original pharmaceutical, with the speed of conversion exhibiting high intra and interpatient reproducibility.
  • a non- enzymatic self cleaving dipeptide/drug complex is provided that comprises a known drug and a dipeptide element covalently bound to the drug through an amide bond.
  • the non-enzymatic self cleaving complex comprises the structure A- B-Q wherein Q is an amine bearing medicinal agent, A is an amino acid or a hydroxyl acid and B is an N-alkylated amino acid that is linked to the medicinal agent through formation of an amide bond between B and an amine of the medicinal agent.
  • the amino acids of the dipeptide are selected such that an intramolecular chemical reaction cleaves A-B from the medicinal agent, producing a diketopiperazine or diketomorpholine and the reconstituted native medicinal agent.
  • a and/or B are selected from non-coding amino acids to inhibit cleavage of the dipeptide from the medicinal agent via an enzymatic mechanism.
  • a and/or B are amino acids in the D-stereoisomer configuration.
  • A is an amino acid in the D stereoisomer configuration and B is an amino acid in the L stereoisomer configuration.
  • A is an amino acid in the L stereoisomer configuration and B is an amino acid in the D stereoisomer configuration.
  • A is an amino acid in the D stereoisomer configuration and B is an amino acid in the D stereoisomer configuration.
  • an injectable depot composition comprising a dipeptide/drug complex having the general structure of A-B-Q and a depot polymer wherein
  • A is an amino acid or a hydroxyl acid
  • B is an N-alkylated amino acid
  • Q is a known drug that comprises an amine, or a derivative of a known drug modified to comprise an amine, wherein one or more depot polymers are linked to the dipeptide/drug complex.
  • the depot polymer is linked to the side chain of A or B, and the dipeptide (A-B) is linked to Q through formation of an amide bond between B and an amine of Q.
  • Q is a medicinal agent.
  • Q is selected from the group of compounds consisting of nuclear hormones, non-glucagon and non- insulin peptide-based hormones, proteins within the class of 4-helix bundle proteins and blood clotting factors.
  • Q is a nuclear hormone or a non- glucagon and non-insulin peptide-based hormone.
  • non-glucagon and non-insulin peptide-based hormones include, but are not limited to, calcitonin (SEQ ID NOs 14-34), parathyroid hormone (PTH; SEQ ID NO: 49), amylin (SEQ ID NOs: 35-47) or pramlitide; (SEQ ID NO: 48), somatostatin (SEQ ID NO: 12 and 13), growth hormone releasing hormone (GHRH; SEQ ID NO: 8), vasopressin (SEQ ID NO: 6), oxytocin (SEQ ID NO: 10), atrial natriuretic factor (ANF; SEQ ID NO: 7), neuropeptide Y (NPY; SEQ ID NO: 9), and pancreatic peptide Y (PYY; SEQ ID NO: 11), or peptides sharing at least 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95 % sequence identity with said non-glucagon and non-insulin peptide-based hormones amino acid sequences
  • Q is a compound selected from the group consisting of thyroid hormone, glucocorticoids, estrogens, androgens, vitamin D, calcitonin, parathyroid hormone (PTH), amylin (or pramlitide), growth hormone, somatostatin, growth hormone releasing hormone (GHRH), vasopressin, oxytocin, atrial natriuretic factor (ANF), neuropeptide Y (NPY), pancreatic peptide Y (PYY), leptin, erythropoietin, colony stimulating factors (such as GCSF), interferons (e.g.
  • Q is a compound selected from the group consisting of thyroid hormone, glucocorticoids, estrogens, androgens, vitamin D, calcitonin, parathyroid hormone (PTH) and amylin.
  • Q is a compound selected from the group consisting of thyroid hormone, calcitonin, parathyroid hormone (PTH) and amylin.
  • Q is thyroid hormone.
  • the depot polymer is selected to be of a sufficient size that the complex A-B- Q is effectively sequestered at the site of injection upon injection of the composition, and/or the depot polymer interferes with Q's ability to interact with its natural ligand.
  • one or more depot polymers are covalently linked to A and/or B either directly or indirectly through a linker.
  • one or more depot polymers are non-covalently linked through a high affinity association with A or B (either through direct interaction with A or B or through a linking moiety covalently bound to A or B).
  • Chemical cleavage of A-B from Q produces a diketopiperazine or diketomorpholine and releases the active drug, in a controlled manner over a predetermined duration of time after administration, to distribute systemically in the patient (in those embodiment where the initial complex is initially sequestered) and allows the active drug to interact with its target ligand.
  • an injectable composition comprising a plurality of different dipeptide/drug complexes wherein the dipeptide/drug complexes differ from each other based on the structure of the dipeptide moiety.
  • the dipeptide/drug complexes comprise a compound of the general structure of A-B-Q (as defined immediately above) with a depot polymer linked to A or B, wherein the dipeptide/drug complexes differ from one another based on the substituents of A and/or B.
  • the medicinal agent (Q) is released in a controlled manner over an extended period of time based on the cleavage rates of the individual different complexes.
  • a composition wherein the composition comprises the medicinal agent (Q) in a free form as well as the medicinal agent (Q) covalently bound to the dipeptide element.
  • the administered composition will have an immediate therapeutic effect due to the presence of the active medicinal agent.
  • the dipeptide is cleaved from the A-B-Q complex and releases additional active medicinal agent (Q) at a predetermined time interval after the initial administration of the composition.
  • the depot polymer is selected from biocompatible polymers known to those skilled in the art.
  • the depot polymers typically have a size selected from a range of about 20,000 to 120,000 Daltons. In one embodiment the depot polymer has a size selected from a range of about 40,000 to 100,000 or about 40,000 to 80,000 Daltons. In one embodiment the depot polymer has a size of about 40,000, 50,000, 60,000, 70,000 or 80,000 Daltons.
  • Suitable depot polymers include but are not limited to dextrans, polylactides, polyglycolides, caprolactone-based polymers, poly(caprolactone), polyanhydrides, polyamines, polyesteramides, polyorthoesters, polydioxanones, polyacetals, polyketals, polycarbonates, polyphosphoesters, polyesters, polybutylene terephthalate, polyorthocarbonates, polyphosphazenes, succinates, poly(malic acid), poly(amino acids), polyvinylpyrrolidone, polyethylene glycol, polyhydroxycellulose,
  • the depot polymer is selected from the group consisting of polyethylene glycol, dextran, polylactic acid, polyglycolic acid and a copolymer of lactic acid and glycolic acid, and in one specific embodiment the depot polymer is polyethylene glycol.
  • the depot polymer comprises one or more polyethylene glycol chains linked to the dipeptide element wherein the combined molecular weight of depot polymer(s) is 40,000 to 80,000 Daltons.
  • the depot polymer is linked to the side chain of one of the two amino acids of the dipeptide A-B (or to the side chain of a hydroxyl acid present at position "A" of the dipeptide).
  • the dipeptide A-B comprises a cysteine or lysine residue to provide a reactive group for ease of attachment of the depot polymer.
  • the dipeptide A-B comprises a lysine or cysteine wherein a polyethylene glycol having a molecular weight selected from the range of 40,000 to 80,000 Daltons is covalently linked to the lysine or cysteine side chain.
  • a and/or B are selected to resist cleavage by peptidases present in human serum, including for example dipeptidyl peptidase IV (DPP-IV).
  • DPP-IV dipeptidyl peptidase IV
  • the rate of cleavage of the dipeptide element from the bioactive peptide is not substantially enhanced (e.g., greater than 2X) when the reaction is conducted using physiological conditions in the presence of serum proteases relative to conducting the reaction in the absence of the proteases.
  • the cleavage half-life of A-B from the bioactive peptide in standard PBS under physiological conditions is not more than two, three, four or five fold the cleavage half-life of A-B from the bioactive protein in a solution comprising a DPP-IV protease.
  • the solution comprising a DPP-IV protease is serum, more particularly mammalian serum, including human serum.
  • one of A or B of said A-B dipeptide represents a non- coded amino acid.
  • Q comprises a peptide
  • A, B or the amino acid comprising the amino group of Q to which A-B is linked
  • amino acid "B" is N-alkylated but is not proline.
  • the N-alkyl group of amino acid B is a C C ⁇ alkyl, and in one embodiment is Ci-C6 alkyl.
  • the dipeptide/drug complex may be further modified to comprise a covalently bound acyl group or alkyl group.
  • the acyl group or alkyl group is covalently linked to the side chain of A or B of the dipeptide A-B.
  • dipeptide element (A-B) comprises the structure:
  • R4 and R 8 are independently selected from the group consisting of H, C1-C18 alkyl, C 2 -C 18 alkenyl, (Ci-Cie alkyl)OH, (Ci-Cie alkyl)SH, (C 2 -C 3 alkyl)SCH 3 , (C1-C4 alkyl)CONH 2 , (C1-C4 alkyl)COOH, (C1-C4 alkyl)NH 2 , (C1-C4
  • R 3 is selected from the group consisting of Ci-C 18 alkyl, (Ci-C 18 alkyl)OH, (Ci-Cie alkyl)NH 2 , (Ci-C 18 alkyl)SH, (C 0 -C 4 alkyl)(C 3 -C 6 )cycloalkyl, (C 0 -C 4 alkyl)(C 2 -C 5 heterocyclic), (C 0 -C 4 alkyl)(C 6 -Ci 0 aryl)R 7 , and (C1-C4 alkyl)(C 3 -C 9 heteroaryl) or R 4 and R together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;
  • R 5 is NHR 6 or OH
  • R 6 is H, Ci-C 8 alkyl or R 6 and R 2 together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;
  • R 7 is selected from the group consisting of H and OH, with the proviso that when R 4 and R 3 together with the atoms to which they are attached form a 5 or 6 member heterocyclic ring, then at least one of Ri and R 2 are other than hydrogen.
  • dipeptide element (A-B) comprises the structure::
  • R 2j Ri and R 8 are independently selected from the group consisting of H,
  • C1-C18 alkyl C 2 -Ci 8 alkenyl, (Ci-Ci 8 alkyl)OH, (Ci-Ci 8 alkyl)SH, (C 2 -C 3 alkyl)SCH 3 , (C1-C4 alkyl)CONH 2 , (C1-C4 alkyl)COOH, (C1-C4 alkyl)NH 2 , (C1-C4
  • R 3 is selected from the group consisting of C1-C18 alkyl, (Ci-C ⁇ alkyl)OH, (C1-C18 alkyl)NH 2, (Ci-Cie alkyl)SH, (C 0 -C 4 alkyl)(C 3 -C 6 )cycloalkyl, (C 0 -C 4 alkyl)(C 2 -C 5 heterocyclic), (C 0 -C 4 alkyl)(C 6 -Ci 0 aryl)R 7 , and (C1-C4 alkyl)(C 3 -C 9 heteroaryl) or R 4 and R 3 together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;
  • R 5 is NHR 6 or OH
  • R 6 is H, Q-Cg alkyl or R 6 and Ri together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;
  • R 7 is selected from the group consisting of hydrogen, C1-C18 alkyl, C 2 -Ci8 alkenyl, (C 0 -C 4 alkyl)CONH 2, (C 0 -C 4 alkyl)COOH, (C 0 -C 4 alkyl)NH 2 , (C 0 -C 4 alkyl)OH, and halo;
  • Ri and Rg are independently H or Ci-Cs alkyl
  • R 2 and R 4 are independently selected from the group consisting of H, Ci-Cg alkyl, C 2 -C 8 alkenyl, (C1-C4 alkyl)OH, (C1-C4 alkyl)SH, (C 2 -C 3 alkyl)SCH 3 , (C1-C4 alkyl)CONH 2 , (C1-C4 alkyl)COOH, (C1-C4 alkyl)NH 2 , (C1-C4 alkyl)NHC(NH 2 +) NH 2 , (Co-C 4 alkyl)(C 3 -C 6 cycloalkyl), (C 0 -C 4 alkyl)(C 2 -C 5 heterocyclic), (C 0 -C 4 alkyl)(C 6 -Cio aryl)R 7 , and CH 2 (C3-C9 heteroaryl), or Ri and R 2 together with the atoms to which they are attached form a C3-Ci 2 cycloal
  • R 5 is NHR 6 ;
  • R 6 is H or Ci-Cs alkyl.
  • dipeptide prodrug element comprises the structure of Formula I, wherein
  • Ri and R 8 are independently H or Ci-Cg alkyl
  • R 2 and R 4 are independently selected from the group consisting of H, Ci-Cg alkyl, C 2 -C 8 alkenyl, (C1-C4 alkyl)OH, (C1-C4 alkyl)SH, (C 2 -C 3 alkyl)SCH 3 , (C1-C4 alkyl)CONH 2 , (C1-C4 alkyl)COOH, (C1-C4 alkyl)NH 2 , (C1-C4 alkyl)NHC(NH 2 +) NH 2 , (Co-C 4 alkyl)(C 3 -C 6 cycloalkyl), (C 0 -C 4 alkyl)(C 2 -C 5 heterocyclic), (C 0 -C 4 alkyl)(C 6 -Cio aryl)R 7 , and CH 2 (C 3 -C9 heteroaryl), or Ri and R 2 together with the atoms to which they are attached form a C 3 -C 12 cyclo
  • R 3 is C1-C18 alkyl
  • R 5 is NHR 6 ;
  • R 6 is H or Ci-Cg alkyl
  • R 7 is selected from the group consisting of hydrogen, Q-Cig alkyl, C 2 -Cig alkenyl, (C 0 -C 4 alkyl)CONH 2 , (C 0 -C 4 alkyl)COOH, (C 0 -C 4 alkyl)NH 2 , (C 0 -C 4 alkyl)OH, and halo.
  • R 4 and R3 together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring then at least one of Ri and R 2 are other than hydrogen.
  • both Ri and R 2 are other than hydrogen.
  • the dipeptide element (A-B) is linked to a medicinal agent via a primary amine present on the native drug, or a primary amine introduced into the drug by chemical modification, wherein the substituents of the dipeptide element are selected to provide a dipeptide/drug complex (A-B-Q) wherein the ti/2 of A-B-Q is about 1 hour in standard PBS under physiological conditions.
  • A-B comprises the structure of formula I wherein
  • Ri and R 2 are independently Ci-Cig alkyl or aryl; or Ri and R 2 are linked through -(CH 2 ) P -, wherein p is 2-9;
  • R 3 is Ci-Cig alkyl
  • R 4 and Rg are each hydrogen
  • R5 is an amine
  • prodrugs having a ti /2 of, e.g., about 1 hour comprise a dipeptide prodrug element with la I:
  • Ri and R 2 are independently Ci-Cig alkyl or (C 0 -C 4 alkyl)(C 6 -Cio aryl)R 7 ; or Ri and R 2 are linked through -(CH 2 ) P , wherein p is 2-9; R 3 is C1-C18 alkyl;
  • R 4 and R 8 are each hydrogen
  • R 5 is NH 2 ;
  • R 7 is selected from the group consisting of hydrogen, Q-Qg alkyl, C 2 -Ci 8 alkenyl, (C 0 -C 4 alkyl)CONH 2> (C 0 -C 4 alkyl)COOH, (C 0 -C 4 alkyl)NH 2 , (C 0 -C 4 alkyl)OH, and halo.
  • the substituents of the dipeptide element are selected to provide a complex A-B-Q, wherein the t of A-B-Q is about 6 to about 24 hours in standard PBS under physiological conditions.
  • a dipeptide/medicinal agent complex is provided having the structure A- B-Q and a t of about 6 to about 24 hours in standard PBS under physiological conditions wherein A-B comprises the structure of formula I further wherein
  • Ri and R 2 are independently selected from the group consisting of hydrogen, Ci-Qg alkyl and aryl, or Ri and R 2 are linked through -(CH 2 ) P -, wherein p is 2-9;
  • R 3 is Ci-Ci 8 alkyl or R 3 and R 4 together with the atoms to which they are attached form a 4-12 heterocyclic ring;
  • R 4 and R 8 are independently selected from the group consisting of hydrogen, Ci-Cg alkyl and aryl;
  • R5 is an amine
  • the substituents of the dipeptide element are selected to provide a complex A-B-Q, wherein the ti /2 of A-B-Q is e.g., between about 12 to about 72 hours, or in some embodiments between about 12 to about 48 hours.
  • a dipeptide/medicinal agent complex is provided having the structure A-B-Q and a t 1/2 between about 12 to about 72 hours, or in some embodiments between about 12 to about 48 hours in standard PBS under
  • Ri and R 2 are independently selected from the group consisting of hydrogen, C1-C18 alkyl, (Ci-Ci 8 alkyl)OH, (C1-C4 alkyl)NH 2 , and (C 0 -C 4 alkyl)(C 6 -Ci 0 aryl)R 7 , or Ri and R 2 are linked through (CH 2 ) P , wherein p is 2-9;
  • R3 is C C ⁇ alkyl or R 3 and R 4 together with the atoms to which they are attached form a 4-12 heterocyclic ring;
  • R 4 and R 8 are independently selected from the group consisting of hydrogen, Ci-Ce alkyl and (C 0 -C 4 alkyl)(C 6 -C 10 aryl)R 7 ;
  • R 5 is NH 2 ;
  • R 7 is selected from the group consisting of H, C -Cn alkyl, C 2 -Ci 8 alkenyl, (Co-C 4 alkyl)CONH 2> (C 0 -C 4 alkyl)COOH, (C 0 -C 4 alkyl)NH 2 , (C 0 -C 4 alkyl)OH, and halo;
  • both R and R 2 are not hydrogen and provided that at least one of R 4 or R 8 is hydrogen.
  • a dipeptide/medicinal agent complex having the structure A-B-Q and a t between about 12 to about 72 hours, or in some embodiments between about 12 to about 48 hours in standard PBS under physiological conditions wherei -B comprises the structure:
  • R and R 2 are independently selected from the group consisting of hydrogen, Ci-Cg alkyl and (CrC 4 alkyl)NH 2 , or R and R 2 are linked through (CH 2 ) P , wherein p is 2-9;
  • R 3 is Ci-Cg alkyl or R and R 4 together with the atoms to which they are attached form a 4-6 heterocyclic ring;
  • R 4 is selected from the group consisting of hydrogen and Ci-Cg alkyl; and R 5 is NH 2 ;
  • a dipeptide/medicinal agent complex having the structure A-B-Q and a tm between about 12 to about 72 hours, or in some embodiments between about 12 to about 48 hours in standard PBS under physiological conditions wherein A-B comprises the structure:
  • Ri and R 2 are independently selected from the group consisting of hydrogen, Ci-C 8 alkyl and (C 1 -C4 alkyl)NH 2 ;
  • R 3 is Ci-C 6 alkyl
  • R 4 is hydrogen
  • R 5 is NH 2 ;
  • a dipeptide/medicinal agent complex having the structure A-B-Q and a ti /2 between about 12 to about 72 hours, or in some embodiments between about 12 to about 48 hours in standard PBS under physiological conditions wherein A-B comprises the structure:
  • Ri and R 2 are independently selected from the group consisting of hydrogen and C 1 -C8 alkyl, (Ci-C 4 alkyl)NH 2 , or Ri and R 2 are linked through (CH 2 ) P , wherein p is 2-9;
  • R 3 is Ci-C 8 alkyl
  • R 4 is (Co-C 4 alkyl)(C 6 -Cio aryl)R 7 ;
  • R 5 is NH 2 ;
  • R 7 is selected from the group consisting of hydrogen, Q-Cg alkyl and (Co-C 4 alkyl)OH;
  • the substituents of the dipeptide element are selected to provide a dipeptide/medicinal agent complex (A-B-Q) wherein the ti /2 of A-B-Q is about 72 to about 168 hours in standard PBS under physiological conditions.
  • A-B comprises the structure of formula I wherein
  • Ri is selected from the group consisting of hydrogen, Q-Cg alkyl and aryl; R 3 is C1-C18 alkyl;
  • R 4 and R 8 are each hydrogen
  • R 5 is an amine or N- substituted amine or a hydroxyl
  • Ri is selected from the group consisting of hydrogen, Ci-Cg alkyl and C 5 -C 10 aryl, and in one embodiment Ri is selected from the group consisting of hydrogen, Ci-Cg alkyl and C5-C6 aryl.
  • A-B comprises the structure: wherein
  • Ri is selected from the group consisting of hydrogen, Ci-Cs alkyl and (C 0 -C 4 alkyl)(C 6 -Cio aryl)R 7 ;
  • R 3 is C 1 -C 18 alkyl
  • R 4 and R 8 are each hydrogen
  • Re is H, C 1 -C8 alkyl, or R 6 and Ri together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;
  • R 7 is selected from the group consisting of hydrogen, Q-Cig alkyl, C 2 -Ci 8 alkenyl, (C 0 -C 4 alkyl)CONH 2> (C 0 -C 4 alkyl)COOH, (C 0 -C 4 alkyl)NH 2 , (C 0 -C 4 alkyl)OH, and halo;
  • Ri is alkyl or (C 0 -C 4 alkyl)(C 6 -Cio aryl)R 7 , then Ri and R5 together with the atoms to which they are attached form a 4- 11 heterocyclic ring.
  • Ri is selected from the group consisting of hydrogen, Ci-Cg alkyl and (Co-C 4 alkyl)(Cs-Cio aryl)R 7
  • Ri is selected from the group consisting of hydrogen, Ci-Cg alkyl and (C 0 -C 4 alkyl)(Cs-C 6 aryl)R 7 .
  • the complexes comprising a depot polymer can be administered as an injectable composition to provide a sustained and controlled delivery of a beneficial agent to a subject over a prolonged duration of time.
  • the dipeptide elements disclosed herein can be linked to any medicinal agent via an amide bond linkage and used to treat any disease or condition in accordance with known uses for the parent medicinal agent.
  • the dipeptide/medicinal agent/depot polymer complexes of the present invention can provide a prolonged controlled delivery that is regulated by selection of the dipeptide substituents. In one embodiment the release is controlled over a period from about 6 to about 24 hours, about 48 to about 72 hours, 72 to about 168 hours, or about two weeks to one month after administration.
  • the present disclosure also encompasses the formulation of prodrug derivatives of known medicinal agent useful for treating patients. More particularly, the prodrugs disclosed herein are formulated to enhance the half life of the parent medicinal agent, while allowing for subsequent activation of the prodrug via a non- enzymatic degradation mechanism.
  • the ideal prodrug should be soluble in water at physiological conditions (for example, a pH of 7.2 and 37 °C), and it should be stable in the powder form for long term storage. It should also be immunologically silent and exhibit a low activity relative to the parent drug. Typically the prodrug will exhibit no more than 10% of the activity of the parent drug, in one embodiment the prodrug exhibits less than 10%, less than 5%, about 1%, or less than 1% activity relative to the parent drug.
  • the prodrug when injected in the body, should be quantitatively converted to the active drug within a defined period of time.
  • a known medicinal agents including bioactive peptides and non-peptide drugs such as thyroid hormone, estrogen, testosterone, and glucocorticoids, as well as analogs, derivatives and conjugates of the foregoing.
  • a chemoreversible prodrug derivative of a known drug wherein the drug is modified to have a dipeptide element covalently bound to an active site of the drug via an amide linkage. Covalent attachment of the dipeptide element to an active site of the drug inhibits the activity of the drug until cleavage of the dipeptide element.
  • a prodrug is provided having a non-enzymatic activation half time (tl/2) between 1-720 hrs under physiological conditions.
  • Physiological conditions as disclosed herein are intended to include a temperature of about 35 to 40 °C and a pH of about 7.0 to about 7.4 and more typically include a pH of 7.2 to 7.4 and a temperature of 36 to 38 °C.
  • the rate of cleavage, and thus activation of the prodrug depends on the structure and stereochemistry of the dipeptide element and also on the strength of the dipeptide nucleophile.
  • the prodrugs disclosed herein will ultimately be chemically converted to structures that can be recognized by the native
  • the molecular design disclosed in this application relies upon an intramolecular chemical reaction that is not dependent upon additional chemical additives, or enzymes.
  • the speed of conversion is controlled by the chemical nature of the dipeptide substituent and its cleavage under physiological conditions. Since physiological pH and temperature are tightly regulated within a highly defined range, the speed of conversion from prodrug to drug will exhibit high intra and interpatient reproducibility.
  • prodrugs are provided having half lives of at least 1 hour, and more typically greater than 20 hours. In one embodiment the half life of the prodrug is about 1, 6, 8, 12, 20, 24, 48 or 72 hours. In one embodiment the half life of the prodrug is 100 hours or greater including half lives of up to 168, 336, 504, 672 or 720 hours, wherein the prodrug is converted to the active form at physiological conditions through a non-enzymatic reaction driven by inherent chemical instability.
  • the non-enzymatic activation tl/2 time of the prodrug is between 1-100 hrs, and more typically between 12 and 72 hours, for example, between 12 and 48 hours and between 48 and 72 hours, and in one embodiment the tl/2 is between 24-48 hrs as measured by incubating the prodrug in a phosphate buffer solution (e.g., PBS) at 37 °C and pH of 7.2.
  • a phosphate buffer solution e.g., PBS
  • the non-enzymatic activation ti/2 time of the prodrug is between 1 and 6 hours, for example, about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, or about 6 hours.
  • the non-enzymatic activation t 1/2 time of the prodrug is between 6 and 24 hours.
  • activation of the prodrug occurs after cleavage of an amide bond linked dipeptide, and formation of a diketopiperazine or diketomorpholine, and the active medicinal agent.
  • Specific dipeptides composed of natural, non-coding and/or synthetic amino acids have been identified that facilitate intramolecular
  • Q is a medicinal agent
  • A is an amino acid or a hydroxyl acid
  • B is an N-alkylated amino acid
  • A-B is a dipeptide that is linked to Q through formation of an amide bond between B and an amine of Q, at an active site of Q.
  • the amino acids of the dipeptide A-B are selected such that chemical cleavage of A-B from Q is more than 90% complete within 720 hours after solubilization in a standard PBS solution under physiological conditions.
  • one of A or B represents a non-coded amino acid, or when the dipeptide A-B is linked to Q through an amino acid, the dipeptide A-B is linked to Q through a non-coded amino acid.
  • the dipeptide A-B is linked to Q through an amide bond that does not constitute a peptide bond.
  • the prodrug comprises the dipeptide A-B linked to the active site of a bioactive peptide wherein A, B, or the amino acid comprising the amino group of Q to which A-B is linked is a non-coded amino acid.
  • the prodrug comprises the structure A-B-Q wherein Q is a known drug that comprises an amine, or a derivative of a know drug modified to comprise an amine.
  • Q is selected from the group of compounds consisting of nuclear hormones, non-glucagon and non-insulin peptide-based hormones, proteins within the class of 4-helix bundle proteins and blood clotting factors.
  • Q is a nuclear hormone or a non-glucagon and non- insulin peptide-based hormone.
  • Q is a compound selected from the group consisting of thyroid hormone, glucocorticoids, estrogens, androgens, vitamin D, calcitonin, parathyroid hormone (PTH), amylin, growth hormone, leptin, erythropoietin, colony stimulating factors (such as GCSF), interferons (e.g. alpha and beta isoforms), tissue plasminogen activitors (TPA), and blood clotting factors, such as Factor VII, Factor VIII and Factor IX.
  • Q is a compound selected from the group consisting of thyroid hormone, glucocorticoids, estrogens, androgens, vitamin D, calcitonin, parathyroid hormone (PTH) and amylin.
  • Q is a compound selected from the group consisting of thyroid hormone, calcitonin, parathyroid hormone (PTH) and amylin.
  • Q is thyroid hormone.
  • the dipeptide element (A-B) is designed to cleave based upon an
  • the dipeptide structure is selected to resist cleavage by peptidases present in mammalian sera, including for example dipeptidyl peptidase IV (DPP-IV). Accordingly, in one embodiment the rate of cleavage of the dipeptide element from the bioactive peptide is not substantially enhanced (e.g., greater than 2X) when the reaction is conducted using physiological conditions in the presence of serum proteases relative to conducting the reaction in the absence of the proteases.
  • DPP-IV dipeptidyl peptidase IV
  • the cleavage half-life of A-B from the bioactive peptide in PBS under physiological conditions is not more than two, three, four or five fold the cleavage half-life of A-B from the bioactive protein in a solution comprising a DPP-IV protease.
  • the solution comprising a DPP-IV protease is serum, more particularly mammalian serum, including human serum.
  • a or B of the dipeptide element, or in the case of a bioactive peptide, the amino acid of the bioactive peptide to which A-B is linked is a non-coded amino acid.
  • amino acid "B" is N-alkylated, but is not proline.
  • the N-alkylated group of amino acid B is a Cr Cig alkyl, and in one embodiment is Ci-C6 alkyl.
  • the cleavage half-life of A-B from Q in standard PBS under physiological conditions is not more than two fold the cleavage half-life of A-B from Q in a solution comprising a DPP-IV protease.
  • the solution comprising the DPP- IV protease is serum.
  • an aliphatic amino group of Q i.e., a primary amine
  • Q i.e., a primary amine
  • the dipeptide element is linked to an amino group present in Q, either directly or through a linking moiety.
  • the linking moiety comprises a primary amine bearing acyl group or alkyl group.
  • the dipeptide element can be linked to an amino substituent present on an aryl ring of the peptide, including for example an aromatic amino acid of a bioactive peptide selected from the group consisting of amino-Phe, amino- napthyl alanine, amino tryptophan, amino-phenyl-glycine, amino-homo-Phe, and amino tyrosine.
  • the dipeptide element is linked to the side chain amino group of a lysine amino acid or the aromatic amino group of a 4- aminophenylalanine (substituted for a native phenylalanine or tyrosine residue of the bioactive peptide).
  • the dipeptide element is linked to an amine present on an internal amino acid of a bioactive peptide. In one embodiment is the dipeptide element is linked to a primary amine. In accordance with one embodiment the dipeptide element can be further modified to comprise a hydrophilic moiety. In one embodiment the hydrophilic moiety is a polyethylene glycol chain. In accordance with one embodiment a polyethylene glycol chain of 40k or higher is covalently bound to the side chain of the A or B amino acid of the dipeptide element. In another embodiment the dipeptide element is acylated or alkylated with a fatty acid or bile acid, or salt thereof, e.g.
  • the dipeptide element can be linked to a depot polymer such as dextran or a polyethylene glycol molecule (e.g. having a size of approximately 40,000 to 80,000 daltons) that serves to sequester the prodrug at an injection site until cleavage of the dipeptide releases the active bioactive peptide.
  • a depot polymer such as dextran or a polyethylene glycol molecule (e.g. having a size of approximately 40,000 to 80,000 daltons) that serves to sequester the prodrug at an injection site until cleavage of the dipeptide releases the active bioactive peptide.
  • R 4 and Rg are independently selected from the group consisting of H, C1-C18 alkyl, C 2 -C 18 alkenyl, (Ci-Cie alkyl)OH, (Ci-Cie alkyl)SH, (C 2 -C 3 alkyl)SCH 3 , (C1-C4 alkyl)CONH 2 , (C1-C4 alkyl)COOH, (C1-C4 alkyl)NH 2 , (C1-C4
  • R 3 is selected from the group consisting of C C ⁇ alkyl, (C C ⁇ alkyl)OH, (Ci-Cie alkyl)NH 2, (Ci-Cie alkyl)SH, (C 0 -C 4 alkyl)(C 3 -C 6 )cycloalkyl, (C 0 -C 4 alkyl)(C 2 -C 5 heterocyclic), (C 0 -C 4 alkyl)(C 6 -Ci 0 aryl)R 7 , and (C1-C4 alkyl)(C 3 -C 9 heteroaryl) or R 4 and R together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;
  • R 5 is NHR 6 or OH
  • R 6 is H, Q-Cg alkyl or R 6 and R 2 together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;
  • R 7 is selected from the group consisting of H and OH.
  • R ⁇ R 2> R4 and Rg are independently selected from the group consisting of H, Ci-Qg alkyl, C 2 -C 18 alkenyl, (Ci-C 18 alkyl)OH, (Ci-Cig alkyl)SH, (C 2 -C 3 alkyl)SCH 3 , (C1-C4 alkyl)CONH 2 , (C1-C4 alkyl)COOH, (C1-C4 alkyl)NH 2 , (C1-C4
  • R 3 is selected from the group consisting of C Cig alkyl, (C Cig alkyl)OH, (C1-C18 alkyl)NH 2> (d-Cig alkyl)SH, (C 0 -C 4 alkyl)(C 3 -C 6 )cycloalkyl, (C 0 -C 4 alkyl)(C 2 -C 5 heterocyclic), (C 0 -C 4 alkyl)(C 6 -Ci 0 aryl)R 7 , and (C1-C4 alkyl)(C 3 -C 9 heteroaryl) or R 4 and R 3 together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;
  • R 5 is NHR 6 or OH
  • R 6 is H, Ci-Cg alkyl or R 6 and Ri together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;
  • R 7 is selected from the group consisting of hydrogen, Ci-Cig alkyl, C 2 -Cig alkenyl, (C 0 -C 4 alkyl)CONH 2, (C 0 -C 4 alkyl)COOH, (C 0 -C 4 alkyl)NH 2 , (C 0 -C 4 alkyl)OH, and halo.
  • Rg is H and R5 is NHR 6.
  • the dipeptide element has the structure of Formula I, wherein
  • Ri and Rg are independently H or Ci-Cg alkyl;
  • R 2 and R 4 are independently selected from the group consisting of H, Ci-Cg alkyl, C 2 -C 8 alkenyl, (C 1 -C4 alkyl)OH, (C 1 -C4 alkyl)SH, (C 2 -C 3 alkyl)SCH 3 , (C 1 -C4 alkyl)CONH 2 , (C1-C4 alkyl)COOH, (C1-C4 alkyl)NH 2 , (C1-C4 alkyl)NHC(NH 2 +) NH 2 , (Co-C 4 alkyl)(C 3 -C 6 cycloalkyl), (C 0 -C 4 alkyl)(C 2 -C 5 heterocyclic), (C 0 -C 4 alkyl)(C 6 -Cio aryl)R 7 , and CH 2 (C 3 -C 9 heteroaryl), or Ri and R 2 together with
  • R 5 is NHR 6 ;
  • R 6 is H or Ci-C 8 alkyl.
  • dipeptide prodrug element has the structure of Formula I, wherein
  • Ri and Rg are independently H or Ci-Cg alkyl
  • R 2 and R 4 are independently selected from the group consisting of H, Ci-Cg alkyl, C 2 -C 8 alkenyl, (C 1 -C4 alkyl)OH, (C 1 -C4 alkyl)SH, (C 2 -C 3 alkyl)SCH 3 , (C 1 -C4 alkyl)CONH 2 , (C 1 -C4 alkyl)COOH, (C 1 -C4 alkyl)NH 2 , (C 1 -C4 alkyl)NHC(NH 2 +) NH 2 , (Co-C 4 alkyl)(C 3 -C 6 cycloalkyl), (C 0 -C 4 alkyl)(C 2 -C 5 heterocyclic), (C 0 -C 4 alkyl)(C 6 -Cio aryl)R 7 , and CH 2 (C 3 -C 9 heteroaryl), or Ri and R 2 together with the atoms to which they are attached form a C 3
  • R 3 is C 1 -C18 alkyl
  • R 5 is NHR 6 ;
  • R 6 is H or Ci-C8 alkyl
  • R 7 is selected from the group consisting of hydrogen, Q-Cig alkyl, C 2 -Cig alkenyl, (C 0 -C 4 alkyl)CONH 2> (C 0 -C 4 alkyl)COOH, (C 0 -C 4 alkyl)NH 2 , (C 0 -C 4 alkyl)OH, and halo.
  • the half life of the prodrug formed in accordance with the present disclosure is determined by the substituents of the dipeptide element and the site on the drug to which it is attached.
  • the prodrug may comprise a dipeptide element linked through an aliphatic amino group of the drug.
  • prodrugs having a t of 1 hour comprise a dipeptide element with the structure:
  • Ri and R 2 are independently Ci-Cis alkyl or aryl; or Ri and R 2 are linked through -(CH 2 ) p , wherein p is 2-9;
  • R 3 is C1-C18 alkyl
  • R 4 and Rg are each hydrogen
  • R5 is an amine
  • prodrugs comprising a dipeptide element linked through an aliphatic amino group of the drug and having a t , e.g., of about 1 hour have the structure:
  • Ri and R 2 are independently Ci-C% alkyl or (C 0 -C4 alkyl)(C 6 -Cio aryl)R 7 ; or Ri and R 2 are linked through -(CH 2 ) P -, wherein p is 2-9;
  • R 3 is C 1 -C 18 alkyl
  • R 4 and R 8 are each hydrogen
  • R 5 is NH 2 ;
  • R 7 is selected from the group consisting of hydrogen, C 1 -C18 alkyl, C 2 -Ci 8 alkenyl, (C 0 -C 4 alkyl)CONH 2> (C 0 -C 4 alkyl)COOH, (C 0 -C 4 alkyl)NH 2 , (C 0 -C 4 alkyl)OH, and halo.
  • prodrugs having the dipeptide element linked through an aliphatic amino group of the drug and having a tl/2 between about 6 to about 24 hours comprise a dipeptide element with the structure:
  • Ri and R 2 are independently selected from the group consisting of hydrogen, C 1 -C 18 alkyl and aryl, or Ri and R 2 are linked through (CH 2 ) P , wherein p is 2-9;
  • R 3 is C 1 -C 18 alkyl or R 3 and R 4 together with the atoms to which they are attached form a 4-12 heterocyclic ring;
  • R 4 and R 8 are independently selected from the group consisting of hydrogen, Ci-Cg alkyl and aryl; and R5 is an amine;
  • both R and R 2 are not hydrogen and provided that one of R 4 or Rg is hydrogen.
  • prodrugs having the dipeptide element linked through an aliphatic amino group of the drug and having a tl/2 between about 12 to about 72 hours, or in some embodiments between about 12 to about 48 hours comprise a dipeptide element with the structure:
  • R and R 2 are independently selected from the group consisting of hydrogen, C1-C18 alkyl, (Ci-Cie alkyl)OH, (C1-C4 alkyl)NH 2 , and (C 0 -C 4 alkyl)(C 6 -C 10 aryl)R 7 , or Ri and R 2 are linked through (CH 2 ) P , wherein p is 2-9;
  • R 3 is C Cig alkyl or R 3 and R 4 together with the atoms to which they are attached form a 4-12 heterocyclic ring;
  • R 4 and Rg are independently selected from the group consisting of hydrogen, Ci-Cg alkyl and (C 0 -C 4 alkyl)(C 6 -Ci 0 aryl)R 7 ;
  • R 5 is NH 2 ;
  • R 7 is selected from the group consisting of H, C Qg alkyl, C 2 -Cig alkenyl, (Co-C 4 alkyl)CONH 2, (C 0 -C 4 alkyl)COOH, (C 0 -C 4 alkyl)NH 2 , (C 0 -C 4 alkyl)OH, and halo;
  • prodrugs having the dipeptide element linked through an aliphatic amino group of the drug and having a tl/2 between about 12 to about 72 hours, or in some embodiments between about 12 to about 48 hours comprise a dipeptide element with the structure:
  • Ri and R 2 are independently selected from the group consisting of hydrogen, Ci-Cg alkyl and (Ci-C 4 alkyl)NH 2 , or Ri and R 2 are linked through (CH 2 ) P , wherein p is 2-9;
  • R 3 is Ci-Cg alkyl or R and R 4 together with the atoms to which they are attached form a 4-6 heterocyclic ring;
  • R 4 is selected from the group consisting of hydrogen and Ci-Cg alkyl; and R 5 is NH 2 ;
  • prodrugs having the dipeptide element linked through an aliphatic amino group of the drug and having a tl/2 between about 12 to about 72 hours, or in some embodiments between about 12 to about 48 hours comprise a dipeptide element with the structure:
  • Ri and R 2 are independently selected from the group consisting of hydrogen, Ci-Ce alkyl and (Ci-C 4 alkyl)NH 2 ;
  • R 3 is Ci-C 6 alkyl
  • R 4 is hydrogen
  • R 5 is NH 2 ;
  • prodrugs having the dipeptide element linked through an aliphatic amino group of the drug and having a tl/2 between about 12 to about 72 hours, or in some embodiments between about 12 to about 48 hours comprise a dipeptide element with the structure:
  • Ri and R 2 are independently selected from the group consisting of hydrogen and Ci-C8 alkyl, (CrC 4 alkyl)NH 2 , or Ri and R 2 are linked through (CH 2 ) P , wherein p is 2-9;
  • R 3 is Ci-Cg alkyl
  • R 4 is (Co-C 4 alkyl)(C 6 -Cio aryl)R 7 ;
  • R 5 is NH 2 ;
  • R 7 is selected from the group consisting of hydrogen, Ci-Cg alkyl and (C 0 -C 4 alkyl)OH;
  • a prodrug having the dipeptide element linked through an aliphatic amino group of the drug and having a tl/2 of about 72 to about 168 hours wherein the dipeptide element has the structure:
  • Ri is selected from the group consisting of hydrogen, Ci-Cg alkyl and aryl;
  • R 3 is Ci-Cie alkyl
  • R 4 and Rg are each hydrogen
  • R5 is an amine or N-substituted amine or a hydroxyl
  • Ri is alkyl or aryl, then Ri and R5 together with the atoms to which they are attached form a 4-11 heterocyclic ring.
  • a prodrug having the dipeptide element linked through an aliphatic amino group of the drug and having a tl/2 of about 72 to about 168 hours is provided wherein the dipepti ture:
  • Ri is selected from the group consisting of hydrogen, Ci-Cig alkyl and (Co-C 4 alkyl)(C 6 -Ci 0 aryl)R 7 ;
  • R 3 is Ci-Cig alkyl
  • R 4 and R 8 are each hydrogen; R 5 is NHR 6 or OH;
  • R 6 is H or Ci-Cg alkyl, or R 6 and Ri together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;
  • R 7 is selected from the group consisting of hydrogen, Q-Cig alkyl, C 2 -Cig alkenyl, (C 0 -C 4 alkyl)CONH 2> (C 0 -C 4 alkyl)COOH, (C 0 -C 4 alkyl)NH 2 , (C 0 -C 4 alkyl)OH, and halo;
  • Ri and R 2 are both independently an alkyl or (C 0 -C 4 alkyl)(C 6 -Cio aryl)R 7 , either Ri or R 2 is linked through (CH 2 ) P to R 5 , wherein p is 2-9.
  • the dipeptide element is linked to a side chain amine of an internal amino acid of a bioactive peptide.
  • prodrugs having a t 2 of about 1 hour have the structu
  • Ri and R 2 are independently Ci-Cg alkyl or aryl; or Ri and R 2 are linked through (CH 2 ) P , wherein p is 2-9;
  • R 3 is C1-C18 alkyl
  • R 4 and Rg are each hydrogen; and R 5 is an amine.
  • the dipeptide element linked to a side chain amine of an internal amino acid of a bioactive peptide and having a t 2 , e.g., of about 1 hour has the structure:
  • Ri and R 2 are independently Ci-Cg alkyl or (Co-C 4 alkyl)(C 6 -Cio aryl)R 7 ; or Ri and R 2 are linked through -(CH 2 ) P -, wherein p is 2-9;
  • R 3 is C 1 -C18 alkyl
  • R 4 and Rg are each hydrogen
  • R 5 is NH 2 ; and R 7 is selected from the group consisting of hydrogen, Q-Cig alkyl, C 2 -Cig alkenyl, (C 0 -C 4 alkyl)CONH 2> (C 0 -C 4 alkyl)COOH, (C 0 -C 4 alkyl)NH 2 , (C 0 -C 4 alkyl)OH, and halo.
  • prodrugs having a tl/2 between about 6 to about 24 hours and having the dipeptide element linked to an internal amino acid side chain comprise a dipeptide element w
  • R and R 2 are independently selected from the group consisting of hydrogen, C C 8 alkyl and aryl, or Ri and R 2 are linked through -(CH 2 ) P , wherein p is 2-9;
  • R 3 is Ci-Cig alkyl or R and R 4 together with the atoms to which they are attached form a 4-12 heterocyclic ring;
  • R 4 and Rg are independently Ci-Cig alkyl or aryl
  • R 5 is an amine or N-substituted amine
  • prodrugs having a ti /2 e.g., between about 12 to about 72 hours, or in some embodiments between about 12 to about 48 hours, and having the dipeptide prodrug element linked to a internal amino acid side chain of a bioactive peptide comprises a dipeptide p structure:
  • Ri and R 2 are independently selected from the group consisting of hydrogen, C Cg alkyl, and (C 0 -C 4 alkyl)(C 6 -Cio aryl)R 7 , or Ri and R 2 are linked through - (CH 2 ) P -, wherein p is 2-9;
  • R 3 is Ci-Cig alkyl or R and R 4 together with the atoms to which they are attached form a 4-12 heterocyclic ring;
  • R 4 and Rg are independently hydrogen, Ci-Cig alkyl or (C 0 -C 4 alkyl)(C 6 -Cio aryl)R 7 ;
  • R 5 is NHR 6 ;
  • R 6 is H or Ci-Cg alkyl, or R 6 and R 2 together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;
  • R 7 is selected from the group consisting of hydrogen, Q-Cig alkyl, C 2 -Ci 8 alkenyl, (C 0 -C 4 alkyl)CONH 2> (C 0 -C 4 alkyl)COOH, (C 0 -C 4 alkyl)NH 2 , (C 0 -C 4 alkyl)OH, and halo;
  • prodrugs having a t 1/2 e.g., between about 12 to about 72 hours, or in some embodiments between about 12 to about 48 hours, and having the dipeptide prodrug element linked to a internal amino acid side chain of a bioactive peptide comprises a dipeptide prodrug element with the structure:
  • Ri and R 2 are independently selected from the group consisting of hydrogen, Ci-Cg alkyl and (Ci-C 4 alkyl)NH 2 , or Ri and R 2 are linked through (CH 2 ) P , wherein p is 2-9;
  • R 3 is Ci-Cg alkyl or R and R 4 together with the atoms to which they are attached form a 4-6 heterocyclic ring;
  • R 4 is selected from the group consisting of hydrogen and Ci-Cg alkyl; and R 5 is NH 2 ;
  • prodrugs having a ti /2 e.g., between about 12 to about 72 hours, or in some embodiments between about 12 to about 48 hours, and having the dipeptide prodrug element linked to a internal amino acid side chain of a bioactive peptide comprises a dipeptide prodrug element with the structure:
  • Ri and R 2 are independently selected from the group consisting of hydi Ci-Cg alkyl and (Ci-C 4 alkyl)NH 2 ;
  • R 3 is Ci-C 6 alkyl;
  • R 4 is hydrogen
  • R 5 is NH 2 ;
  • prodrugs having a t 1/2 e.g., between about 12 to about 72 hours, or in some embodiments between about 12 to about 48 hours, and having the dipeptide prodrug element linked to a internal amino acid side chain of a bioactive peptide comprises a dipeptide prodrug element with the structure:
  • Ri and R 2 are independently selected from the group consisting of hydrogen and C C 8 alkyl, (CrC 4 alkyl)NH 2 , or R and R 2 are linked through (CH 2 ) P , wherein p is 2-9;
  • R 3 is Ci-C 8 alkyl
  • R 4 is (Co-C 4 alkyl)(C 6 -Cio aryl)R 7 ;
  • R 5 is NH 2 ;
  • R 7 is selected from the group consisting of hydrogen, Q-Cg alkyl and (Co-C 4 alkyl)OH;
  • a prodrug having a tl/2 of about 72 to about 168 hours and having the dipeptide element linked to an internal amino acid side chain is provided wherein the dipeptide element has the s
  • R and R 2 are independently selected from the group consisting of hydrogen, C C ⁇ alkyl and aryl;
  • R 3 is C 1 -C18 alkyl
  • R 4 and R 8 are each hydrogen
  • R5 is an amine or N-substituted amine or a hydroxyl; with the proviso that, if Ri and R 2 are both independently an alkyl or aryl, either Ri or R 2 is linked through (CH2) P to R5, wherein p is 2-9.
  • a prodrug having a t e.g., of about 72 to about 168 hours and having the dipeptide prodrug element linked to an internal amino acid side chain is provided wherein the dipeptide prodrug element has the structure:
  • Ri and R 2 are independently selected from the group consisting of hydrogen, C1-C18 alkyl and (C0-C4 alkyl)(C6-Cio aryl)R 7 ;
  • R 3 is C1-C18 alkyl
  • R 4 and R 8 are each hydrogen
  • R 5 is NHR 6 or OH
  • R 6 is H or Ci-C8 alkyl, or R 6 and Ri together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;
  • R 7 is selected from the group consisting of hydrogen, Q-Cig alkyl, C 2 -Cig alkenyl, (C 0 -C 4 alkyl)CONH 2, (C 0 -C 4 alkyl)COOH, (C 0 -C 4 alkyl)NH 2 , (C 0 -C 4 alkyl)OH, and halo;
  • Ri and R 2 are both independently an alkyl or (Co-C 4 alkyl)(C 6 -Cio aryl)R 7 , either Ri or R 2 is linked through (CH 2 ) P to R5, wherein p is 2-9.
  • n is an integer selected from 1 to 4. In one embodiment n is 3 or 4 and in one embodiment the internal amino acid is lysine.
  • the dipeptide element is linked to the bioactive peptide via an amine substituent of an aryl group present in the bioactive peptide. In one embodiment the amino group substituent is a primary amine.
  • prodrugs having a t 1/2 of about 1 hour have a dipeptide structure of:
  • R and R 2 are independently C C ⁇ alkyl or aryl
  • R3 is C C ⁇ alkyl or R 3 and R 4 together with the atoms to which they are attached form a 4-12 heterocyclic ring;
  • R 4 and R 8 are independently selected from the group consisting of hydrogen, C C ⁇ alkyl and aryl; and R5 is an amine or a hydroxyl.
  • prodrugs having a t 1/2 of about 1 hour have a dipeptide structure of:
  • Ri and R 2 are independently C -Cn alkyl or (Co-C 4 alkylXCe-Cio aryl)R 7 ;
  • R 3 is C C ⁇ alkyl or R 3 and R 4 together with the atoms to which they are attached form a 4-12 heterocyclic ring;
  • R 4 and R 8 are independently selected from the group consisting of hydrogen, C1-C18 alkyl and (C 0 -C 4 alkyl)(C 6 -C 10 aryl)R 7 ;
  • R 5 is NH 2 or OH
  • R 7 is selected from the group consisting of hydrogen, Q-Cig alkyl, C 2 -Ci 8 alkenyl, (C 0 -C 4 alkyl)CONH 2> (C 0 -C 4 alkyl)COOH, (C 0 -C 4 alkyl)NH 2 , (C 0 -C 4 alkyl)OH, and halo.
  • prodrugs having the dipeptide element linked to the medicinal agent via an amine substituent of an aryl group present in the medicinal agent, and having a tl/2 of about 6 to about 24 hours are provided wherein the dipeptide comprises a structure of:
  • Ri is selected from the group consisting of hydrogen, Q-Cig alkyl and aryl, or Ri and R 2 are linked through -(CH 2 ) P , wherein p is 2-9;
  • R 3 is Ci-Cig alkyl or R and R 4 together with the atoms to which they are attached form a 4-6 heterocyclic ring;
  • R 4 and Rg are independently selected from the group consisting of hydrogen, Ci-Qg alkyl and aryl; and R5 is an amine or N-substituted amine.
  • prodrugs having the dipeptide prodrug element linked via an aromatic amino acid and having a ti /2 , e.g., of about 6 to about 24 hours are provided wherein the dipeptide comprises a structure of:
  • Ri is selected from the group consisting of hydrogen, Q-Cig alkyl, (Ci-Cig alkyl)OH, (C1-C4 alkyl)NH 2 , and (C 0 -C 4 alkyl)(C 6 -Ci 0 aryl)R 7 ;
  • R 3 is Ci-Cig alkyl or R 3 and R 4 together with the atoms to which they are attached form a 4-6 heterocyclic ring;
  • R 4 and R 8 are independently selected from the group consisting of hydrogen, Ci-Cig alkyl and (C 0 -C 4 alkyl)(C 6 -Ci 0 aryl)R 7 ;
  • R 5 is NHR 6 ;
  • R 6 is H, C Cg alkyl, or R 6 and Ri together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;
  • R 7 is selected from the group consisting of hydrogen, Q-Cig alkyl, C 2 -Cig alkenyl, (C 0 -C 4 alkyl)CONH 2, (C 0 -C 4 alkyl)COOH, (C 0 -C 4 alkyl)NH 2 , (C 0 -C 4 alkyl)OH, and halo.
  • prodrugs having the dipeptide element linked to the medicinal agent via an amine substituent of an aryl group present in the medicinal agent, and having a tl/2 of about 72 to about 168 hours are provided wherein the dipeptide comprises a structure of:
  • Ri and R 2 are independently selected from the group consisting of hydrogen, Ci-C 8 alkyl and aryl;
  • R 3 is CrC 18 ;
  • R 4 and R 8 are each hydrogen
  • Rs is selected from the group consisting of amine, N-substituted amine and hydroxyl.
  • prodrugs having the dipeptide prodrug element linked via an aromatic amino acid and having a ti /2 , e.g., of about 72 to about 168 hours are provided wherein the dipeptide comprises a structure of:
  • Ri and R 2 are independently selected from the group consisting of hydrogen, Ci-C 8 alkyl, (C1-C4 alkyl)COOH, and (C 0 -C 4 alkyl)(C 6 -C 10 aryl)R 7 , or Ri and R5 together with the atoms to which they are attached form a 4- 11 heterocyclic ring;
  • R 3 is Ci-C 18 alkyl or R and R 4 together with the atoms to which they are attached form a 4-6 heterocyclic ring;
  • R 4 is hydrogen or forms a 4-6 heterocyclic ring with R 3 ;
  • R 8 is hydrogen
  • R 5 is NHR 6 or OH
  • R 6 is H or Ci-C 8 alkyl, or R 6 and Ri together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring; and R 7 is selected from the group consisting of hydrogen, Q-Cig alkyl, C 2 -Cig alkenyl, (C 0 -C 4 alkyl)CONH 2> (C 0 -C 4 alkyl)COOH, (C 0 -C 4 alkyl)NH 2 , (C 0 -C 4 alkyl)OH, and halo.
  • the dipeptide element is linked to a bioactive peptide via an amine present on an aryl group of an aromatic amino acid present in the bioactive peptide.
  • the aromatic amino acid is an internal amino acid of the medicinal agent, however the aromatic amino acid can also be the N-terminal amino acid.
  • the aromatic amino acid is selected from the group consisting of amino-Phe, amino-napthyl alanine, amino tryptophan, amino-phenyl- glycine, amino-homo-Phe, and amino tyrosine.
  • the primary amine that forms an amide bond with the dipeptide element is in the para-position on the aryl group.
  • the aromatic amine comprises the structure of Formula III:
  • n is an integer from 1 to 3.
  • Ri is selected from the group consisting of H and Ci-Cg alkyl
  • R 2 and R 4 are independently selected from the group consisting of H, Q-Cg alkyl, C 2 -C 8 alkenyl, (Ci-C 4 alkyl)OH, (Ci-C 4 alkyl)SH, (C 2 -C 3 alkyl)SCH 3 , (Ci-C 4 alkyl)CONH 2 , (Ci-C 4 alkyl)COOH, (Ci-C 4 alkyl)NH 2 , (Ci-C 4 alkyl)NHC(NH 2 + ) NH 2 , (Co-C 4 alkyl)(C 3 -C 6 cycloalkyl), (C 0 -C 4 alkyl)(C 6 -Ci 0 aryl)R 7 , CH 2 (C 5 -C 9 heteroaryl), or Ri and R 2 together with the atoms to which they are attached form a C 3 -C 6 cycloalkyl; R 3 is selected from the group consisting of alkyl, (C 3
  • R 5 is NHR 6 or OH
  • R 6 is H, or R 6 and R 2 together with the atoms to which they are attached form a 5 or 6 member heterocyclic ring;
  • R 7 is selected from the group consisting of H and OH.
  • R is H or Ci-Cg alkyl
  • R 2 is selected from the group consisting of H, C C 6 alkyl, CH 2 OH, (Ci-C 4 alkyl)NH 2 , (C 3 -C 6 cycloalkyl) and CH 2 (C 6 aryl)R 7 or R 6 and R 2 together with the atoms to which they are attached form a 5 member heterocyclic ring.
  • R 3 is C C 6 alkyl
  • R 4 is selected from the group consisting of H, CrC 4 alkyl, (C 3 - C 6 )cycloalkyl, (C1-C4 alkyl)OH, (C1-C4 alkyl)SH and (C 0 -C 4 alkyl)(C 6 aryl)R 7 , or R 3 and R 4 together with the atoms to which they are attached form a 5 member heterocyclic ring.
  • R is CH
  • R5 is NHR 6
  • R 3 and R ⁇ together with the atoms to which they are attached form a 5 member heterocyclic ring and R5 is NHR 6 .
  • R is selected from the group consisting of H and Ci-Cg alkyl
  • R 2 and R 4 are independently selected from the group consisting of H, C Cg alkyl, C 2 -C 8 alkenyl, (C 1 -C4 alkyl)OH, (C 1 -C4 alkyl)SH, (C 2 -C 3 alkyl)SCH 3 , (C 1 -C4 alkyl)CONH 2 , (C 1 -C4 alkyl)COOH, (C 1 -C4 alkyl)NH 2 , (C 1 -C4 alkyl)NHC(NH 2 + ) NH 2 , (Co-C 4 alkyl)(C 3 -C 6 cycloalkyl), (C 0 -C 4 alkyl)(C 6 -C 10 aryl)R 7 , CH 2 (C 5 -C 9 heteroaryl), or Ri and R 2 together with the atoms to which they are attached form a C 3 -C 6 cycloalkyl;
  • R 3 is selected from the group consisting of Ci-Cg alkyl, (C 3 -C 6 )cycloalkyl or R 4 and R 3 together with the atoms to which they are attached form a 5 or 6 member heterocyclic ring;
  • R 5 is NHR 6 or OH
  • R 6 is H, or R 6 and R 2 together with the atoms to which they are attached form a 5 or 6 member heterocyclic ring
  • R 7 is selected from the group consisting of hydrogen, Q-C ⁇ alkyl, C 2 -Ci8 alkenyl, (C 0 -C 4 alkyl)CONH 2 , (C 0 -C 4 alkyl)COOH, (C 0 -C 4 alkyl)NH 2 , (C 0 -C 4 alkyl)OH, and halo.
  • Ri is H or Ci-Cg alkyl
  • R 2 is selected from the group consisting of H, Ci-C 6 alkyl, CH 2 OH, (C C 4 alkyl)NH 2 , (C 3 -C 6 cycloalkyl) and CH 2 (C6 aryl)R 7 or R 6 and R 2 together with the atoms to which they are attached form a 5 member heterocyclic ring
  • R 3 is CrC 6 alkyl
  • R 4 is selected from the group consisting of H, C C 4 alkyl, (C 3 -C 6 )cycloalkyl, (C C 4 alkyl)OH, (C C 4 alkyl)SH and (Co-C 4 alkyl)(C 6 aryl)R 7 , or R and R 4 together with the atoms to which they are attached form a 5 member heterocyclic ring.
  • R 3 is CH 3
  • R 5 is NHR 6
  • the following compounds are provided as examples of compounds that can be combined with the prodrug elements disclosed herein to form prodrug derivatives or sequestered complexes of the known drugs and bioactive peptides.
  • Glucocorticoids a class of corticosteroids, are endogenous hormones with profound effects on the immune system and multiple organ systems. They suppress a variety of immune and inflammatory functions by inhibition of inflammatory cytokines such as IL-1, IL-2, IL-6, and TNF, inhibition of arachidonic acid metabolites including prostaglandins and leukotrienes, depletion of T-lymphocytes, and reduction of the expression of adhesion molecules on endothelial cells (P. J. Barnes, Clin. Sci., 1998, 94, pp. 557-572; P. J. Barnes et al., Trends Pharmacol. Sci., 1993, 14, pp. 436-441). In addition to these effects, glucocorticoids stimulate glucose production in the liver and catabolism of proteins, play a role in electrolyte and water balance, reduce calcium absorption, and inhibit osteoblast function.
  • glucocorticoids The effects of glucocorticoids are mediated at the cellular level by the glucocorticoid receptor (R. H. Oakley and J. Cidlowski, Glucocorticoids, N. J.
  • the glucocorticoid receptor is a member of a class of structurally related intracellular receptors that when coupled with a ligand can function as a transcription factor that affects gene expression (R. M. Evans, Science, 1988, 240, pp. 889-895).
  • Other members of the family of steroid receptors include the mineralocorticoid,
  • glucocorticoids The anti-inflammatory and immune suppressive activities of endogenous glucocorticoids have stimulated the development of synthetic glucocorticoid derivatives including dexamethasone, prednisone, and prednisolone (L. Parente, Glucocorticoids, N. J. Goulding and R. J. Flowers (eds.), Boston: Birkhauser, 2001, pp. 35-54).
  • rheumatic diseases such as rheumatoid arthritis, juvenile arthritis, and ankylosing spondylitis
  • dermatological diseases including psoriasis and pemphigus
  • allergic disorders including allergic rhinitis, atopic dermatitis, and contact dermatitis
  • pulmonary conditions including asthma and chronic obstructive pulmonary disease (COPD)
  • COPD chronic obstructive pulmonary disease
  • Crohn's disease ulcerative colitis
  • systemic lupus erythematosus autoimmune chronic active hepatitis
  • osteoarthritis tendonitis
  • bursitis J.
  • PCT International Publication No. WO 99/33786 discloses triphenylpropanamide compounds with potential use in treating inflammatory diseases.
  • PCT International Publication No. WO 00/66522 describes non-steroidal compounds as selective modulators of the glucocorticoid receptor potentially useful in treating metabolic and inflammatory diseases.
  • PCT International Publication No. WO 99/41256 describes tetracyclic modulators of the glucocorticoid receptor potentially useful in treating immune, autoimmune, and inflammatory diseases.
  • U.S. Pat. No. 5,688,810 describes various non-steroidal compounds as modulators of glucocorticoid and other steroid receptors.
  • PCT International Publication No. WO 99/33786 discloses triphenylpropanamide compounds with potential use in treating inflammatory diseases.
  • PCT International Publication No. WO 00/66522 describes non-steroidal compounds as selective modulators of the glucocorticoid receptor potentially useful in treating metabolic and inflammatory diseases.
  • WO 00/32584 discloses non-steroidal compounds having antiinflammatory activity with dissociation between anti-inflammatory and metabolic effects.
  • PCT International Publication No. WO 98/54159 describes non-steroidal cyclically substituted acylanilides with mixed gestagen and androgen activity.
  • U.S. Pat. No. 4,880,839 describes acylanilides having progestational activity and EP 253503 discloses acylanilides with antiandrogenic properties.
  • PCT International Publication No. WO 97/27852 describes amides that are inhibitors of farnesyl-protein transferase.
  • a derivative of a glucocorticoid receptor agonist or antagonist comprising the structure A-B-Q.
  • Q is the glucocorticoid receptor agonist or antagonist
  • A is an amino acid or a hydroxyl acid
  • B is an N-alkylated amino acid.
  • a and B together represent the dipeptide element that is linked to Q through formation of an amide bond between A-B and an amine of Q.
  • at least one of A or B is a non-coded amino acid.
  • Q is selected from the group consisting of dexamethasone, prednisone, and prednisolone.
  • the dipeptide element is selected wherein chemical cleavage of A-B from Q is at least about 90% complete within about 1 to about 720 hours in PBS under physiological conditions.
  • amino acids of the dipeptide are selected wherein the cleavage half-life of A-B from Q in PBS under physiological conditions is not more than two to five fold the cleavage half-life of A- B from Q in a solution comprising a DPP-IV protease (including for example, human serum).
  • a DPP-IV protease including for example, human serum.
  • Thyroxine is a thyroid hormone involved in the control of cellular metabolism. Chemically, thyroxine is an iodinated derivative of the amino acid tyrosine. The maintenance of a normal level of thyroxine is important for normal growth and development of children as well as for proper bodily function in the adult. Its absence leads to delayed or arrested development. Hypothyroidism, a condition in which the thyroid gland fails to produce enough thyroxine, leads to a decrease in the general metabolism of all cells, most characteristically measured as a decrease in nucleic acid and protein synthesis, and a slowing down of all major metabolic processes. Conversely, hyperthyroidism is an imbalance of metabolism caused by overproduction of thyroxine.
  • T4 is converted to T3 or to rT3 via removal of an iodine atom from one of the hormonal rings.
  • T3 is the biologically active thyroid hormone, whereas rT3 has no biological activity.
  • Both T3 and T4 are used to treat thyroid hormone deficiency (hypothyroidism). They are both absorbed well by the gut, so can be given orally.
  • a thyroid hormone derivative comprising the structure A-B-Q.
  • Q is the thyroid hormone
  • A is an amino acid or a hydroxyl acid
  • B is an N-alkylated amino acid.
  • a and B together represent the dipeptide element that is linked to Q through formation of an amide bond between A-B and an amine of Q.
  • at least one of A, B, or the amino acid of Q to which A-B is linked, is a non-coded amino acid.
  • Q is selected from the group consisting of thyroxine T4 (3,5,3',5'-tetraiodothyronine), 3,5,3'-triiodo L-thyronine and 3,3',5'-triiodo L- thyronine.
  • the dipeptide element is linked via an amide bond through the primary amine of 3,5,3',5'-tetraiodothyronine or 3,5,3'-triiodo L- thyronine.
  • the dipeptide element is selected wherein chemical cleavage of A-B from Q is at least about 90% complete within about 1 to about 720 hours in PBS under physiological conditions.
  • amino acids of the dipeptide are selected wherein the cleavage half-life of A-B from Q in PBS under physiological conditions is not more than two to five fold the cleavage half-life of A-B from Q in a solution comprising a DPP-IV protease
  • antitumor drugs possess a limited bioavailability due to low chemical stability, a limited oral absorption, or a rapid breakdown in vivo (i.e., by first-pass metabolism).
  • various prodrugs that can be activated into antitumor drugs have been designed. In this case it is preferred if prodrugs are activated relatively slowly in the blood or liver, for example, thereby preventing acute toxic effects due to high peak concentrations of the antitumor drug.
  • An ideal prodrug designed to increase the bioavailability of an antitumor drug is slowly released.
  • the prodrug is targeted to tumor cells by complexing the prodrug with a tumor specific ligand or antibody.
  • the anti-cancer drugs is selected from the group consisting of taxanes, such as paclitaxel or taxotere; camptothecins, such as camptothecin, CPT 11, irinotecan, topotecan or HC1; podophyllotoxins, such as teniposide; vinblastine sulfate;
  • vincristine sulfate vinorelbine tartrate
  • procarbazine HC1 cladribine, leustatin; hydroxyurea
  • gemcitabine HC1 leuprolide acetate
  • thioguanine purinethol
  • anthracyclines such as daunorubicin or doxorubicin (adriamycin);
  • methotrexates p-aminoaniline mustard; cytarabine (ara-C or cytosine arabinoside); etoposide; bleomycin sulfate; actinomycin D; idarubicin HC1; mitomycin; plicamycin; mitoxantrone HC1; pentostatin; streptozocin; L-phenylalanine mustard; carboplatin derivatives; platinol; busulfan; fluconazole; amifostine; leucovorin calcium and octreotide acetate.
  • a known anti-cancer agent derivative comprising the structure A-B-Q.
  • Q is the anti-cancer agent
  • A is an amino acid or a hydroxyl acid
  • B is an N-alkylated amino acid.
  • a and B together represent the dipeptide element that is linked to Q through formation of an amide bond between A-B and an amine of Q.
  • at least one of A, B, or the amino acid of Q to which A-B is linked is a non-coded amino acid.
  • the present invention also provides novel methods of administering compositions and formulations comprising derivatives of known antibiotics.
  • the methods provide compositions of active compounds that, if presented in presently available forms, may result in toxicity to the treated mammal.
  • the formulations and methods of the present invention enable one to administer compounds that previously have not been able to be widely used in particular species due to safety considerations.
  • the methods also enable one to extend the release times of compounds and provide a controlled dose of active compound to the treated patient.
  • a prodrug derivative of a known antibiotic is provided.
  • the antibiotic is selected from the group consisting of oxytetracycline, doxycycline, fluoxetime, roxithromycin, terbinarefine, or metoprolol.
  • Oxytetracycline is a widely used and useful antibiotic for treating various infections in mammals. In particular it is used for treating and preventing respiratory infections in domestic animals. There are significant costs associated with repeated administrations through conventional means.
  • a dipeptide element A-B is covalently linked to an antibiotic, including for example, oxytetracycline, wherein the complex optionally further includes a depot polymer.
  • Additional compounds can be linked to the dipeptide element disclosed herein to form prodrug derivatives or depot derivatives of the compounds.
  • additional compounds include growth factors, both natural and recombinant, as well as peptide fractions of growth factors that bind to receptors on the cell surface (EGF, VEGF, FGF, ILGF-I, ILGF-II, TGF).
  • Prodrug derivatives of interferons both natural or recombinant (including IFN-alpha, beta, and gamma) and interferon agonists; and prodrug derivatives of cytokines, either natural or recombinant, including for example (IL- 1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL- 10, IL- 12, IL- 15, TNF, etc) are also encompassed within the scope of the present invention.
  • any peptide, natural, recombinant, or synthetic that binds to a cell surface receptor can be modified to by linking the dipeptide element disclosed herein to form a prodrug or depot derivative of that peptide.
  • the dipeptide element disclosed herein can be linked to the bioactive peptides disclosed in PCT/US2008/053857 either through the N-terminal amine or to the side chain amino group of a lysine at position 20 or the aromatic amino group of a 4-amino phenylalanine substituted for the amino acid at position 22 of any of the disclosed bioactive peptides.
  • the dipeptide element disclosed herein is linked via an amide bond to the N-terminal amine of a bioactive peptide disclosed in PCT/US2008/053857.
  • a complex comprising a medicinal agent and a dipeptide element, A-B, is provided.
  • the dipeptide A-B comprises the structure:
  • Ri and R 8 are independently H or alkyl;
  • R 2 and R 4 are independently selected from the group consisting of H, alkyl, C 2 -C 8 alkenyl, (C 1 -C4 alkyl)OH, (C 1 -C4 alkyl)SH, (C 2 -C 3 alkyl)SCH 3 , (C 1 -C4 alkyl)CONH 2 , (C1-C4 alkyl)COOH, (C1-C4 alkyl)NH 2 , (C1-C4 alkyl)NHC(NH 2 +) NH 2 , (Co-C 4 alkyl)(C 3 -C 6 cycloalkyl), (C 0 -C 4 alkyl)(C 2 -C 5 heterocyclic), (C 0 -C 4 alkyl)(C 6 -Cio aryl)R 7 , and CH 2 (C 3 -C 9 heteroaryl), or Ri and R 2 together with the atoms to which they are
  • R 5 is NHR 6 ;
  • R 6 is H or Ci-C 8 alkyl.
  • Ri and R 8 are independently H or Ci-Cg alkyl
  • R 2 and R 4 are independently selected from the group consisting of H, Q-Cs alkyl, C 2 -C 8 alkenyl, (C1-C4 alkyl)OH, (C1-C4 alkyl)SH, (C 2 -C 3 alkyl)SCH 3 , (C1-C4 alkyl)CONH 2 , (C 1 -C4 alkyl)COOH, (C 1 -C4 alkyl)NH 2 , (C 1 -C4 alkyl)NHC(NH 2 +) NH 2 , (Co-C 4 alkyl)(C 3 -C 6 cycloalkyl), (C 0 -C 4 alkyl)(C 2 -C 5 heterocyclic), (C 0 -C 4 alkyl)(C 6 -Cio aryl)R 7 , and CH 2 (C 3 -C 9 heteroaryl), or Ri and R 2 together with the atoms to which they are attached form a C 3 -Ci
  • R 3 is C 1 -C18 alkyl
  • R 5 is NHR 6 ;
  • R 6 is H or C 1 -C8 alkyl
  • R 7 is selected from the group consisting of hydrogen, Q-C ⁇ alkyl, C 2 -Ci 8 alkenyl, (Co-C 4 alkyl)CONH 2> (C 0 -C 4 alkyl)COOH, (C 0 -C 4 alkyl)NH 2 , (C 0 -C 4 alkyl)OH, and halo.
  • the dipeptide A-B is linked via an amide bond to an aliphatic amino acid of a compound "Q" as defined herein.
  • Ri and R 2 are independently C 1 -C 18 alkyl or aryl; or Ri and R 2 are linked through -(CH 2 ) P -, wherein p is 2-9;
  • R 3 is C 1 -C 18 alkyl; R 4 and R 8 are each hydrogen; and
  • R5 is an amine
  • the dipeptide A-B comprises the structure:
  • Ri and R 2 are independently Ci-Cig alkyl or (C 0 -C 4 alkyl)(C 6 -Cio aryl)R 7 ; or Ri and R 2 are linked through -(CH 2 ) P , wherein p is 2-9;
  • R 3 is C1-C18 alkyl
  • R 4 and Rg are each hydrogen
  • R 5 is NH 2 ;
  • R 7 is selected from the group consisting of hydrogen, Q-Cig alkyl, C 2 -C 1 g alkenyl, (C 0 -C 4 alkyl)CONH 2> (C 0 -C 4 alkyl)COOH, (C 0 -C 4 alkyl)NH 2 , (C 0 -C 4 alkyl)OH, and halo.
  • Ri and R 2 are independently selected from the group consisting of hydrogen, Ci-Cig alkyl and aryl, or Ri and R 2 are linked through -(CH 2 ) P -, wherein p is 2-9;
  • R 3 is Ci-Cig alkyl or R 3 and R 4 together with the atoms to which they are attached form a 4-12 heterocyclic ring;
  • R 4 and R 8 are independently selected from the group consisting of hydrogen, Ci-Cg alkyl and aryl;
  • R5 is an amine; with the proviso that both Ri and R 2 are not hydrogen and provided that one of R 4 or R 8 is hydrogen.
  • the dipeptide A-B comprises the structure:
  • Ri and R 2 are independently selected from the group consisting of hydrogen, Ci-Cig alkyl, (Ci-Cig alkyl)OH, (C 1 -C4 alkyl)NH 2 , and (C 0 -C 4 alkyl)(C 6 - C 10 aryl)R 7 , or Ri and R 2 are linked through (CH 2 ) P , wherein p is 2-9;
  • R 3 is Ci-Cig alkyl or R 3 and R 4 together with the atoms to which they are attached form a 4-12 heterocyclic ring;
  • R 4 and Rg are independently selected from the group consisting of hydrogen, Ci-Ce alkyl and (C 0 -C 4 alkyl)(C 6 -C 10 aryl)R 7 ;
  • R 5 is NH 2 ;
  • R 7 is selected from the group consisting of H, Ci-Cig alkyl, C 2 -Cig alkenyl, (Co-C 4 alkyl)CONH 2, (C 0 -C 4 alkyl)COOH, (C 0 -C 4 alkyl)NH 2 , (C 0 -C 4 alkyl)OH, and halo;
  • Ri is selected from the group consisting of hydrogen, Q-Cg alkyl and aryl;
  • R 3 is C1-C18 alkyl
  • R 4 and R 8 are each hydrogen
  • R 5 is an amine or N-substituted amine or a hydroxyl
  • Ri is alkyl or aryl
  • R 5 together with the atoms to which they are attached form a 4-11 heterocyclic ring.
  • a dipeptide element is provided:
  • Ri is selected from the group consisting of hydrogen, Q-Cig alkyl and (Co-C 4 alkyl)(C 6 -Ci 0 aryl)R 7 ;
  • R 3 is C1-C18 alkyl
  • R 4 and R 8 are each hydrogen
  • R 5 is NHR 6 or OH
  • R 6 is H or Ci-C8 alkyl, or R 6 and Ri together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;
  • R 7 is selected from the group consisting of hydrogen, Q-Cig alkyl, C 2 -Cig alkenyl, (C 0 -C 4 alkyl)CONH 2> (C 0 -C 4 alkyl)COOH, (C 0 -C 4 alkyl)NH 2 , (C 0 -C 4 alkyl)OH, and halo;
  • Ri is alkyl or (C 0 -C 4 alkyl)(C6-Cio aryl)R 7 , Ri is linked through (CH 2 ) P to R5, wherein p is 2-9.
  • a dipeptide element is provided:
  • Ri and R 2 are independently selected from the group consisting of hydrogen, Ci-Cg alkyl and (Ci-C 4 alkyl)NH 2 , or Ri and R 2 are linked through (CH 2 ) P , wherein p is 2-9;
  • R 3 is Ci-Cg alkyl or R 3 and R 4 together with the atoms to which they are attached form a 4-6 heterocyclic ring;
  • R 4 is selected from the group consisting of hydrogen and Ci-Cg alkyl; and R 5 is NH 2 ;
  • a dipeptide element is provided:
  • Ri and R 2 are independently selected from the group consisting of hydrogen, Ci-C 8 alkyl and (C C 4 alkyl)NH 2 ;
  • R 3 is Ci-C 6 alkyl
  • R 4 is hydrogen
  • R 5 is NH 2 ;
  • Ri and R 2 are independently selected from the group consisting of hydrogen and Ci-Cg alkyl, (Ci-C 4 alkyl)NH 2 , or Ri and R 2 are linked through (CH 2 ) P , wherein p is 2-9;
  • R 3 is Ci-Cg alkyl
  • R 4 is (Co-C 4 alkyl)(C 6 -C 10 aryl)R 7 ;
  • R 5 is NH 2 ;
  • R 7 is selected from the group consisting of hydrogen, Q-Cg alkyl and (C 0 -C 4 alkyl)OH;
  • the dipeptide element (A-B) is linked via an amide bond to an amine substituent on an aryl group of Q of the complex A-B-Q.
  • the dipeptide element comprises the structure of formula I linked via an amide bond to an amine substituent on an aryl
  • Ri and R 2 are independently Ci-Cig alkyl or aryl
  • R 3 is Ci-Ci 8 alkyl or R 3 and R 4 together with the atoms to which they are attached form a 4-12 heterocyclic ring;
  • R 4 and R 8 are independently selected from the group consisting of hydrogen, Ci-Cig alkyl and aryl;
  • R 5 is an amine or a hydroxyl.
  • the dipeptide element comprises the structure of formula I linked via an amide bond to an amine substituent on an aryl
  • Ri and R 2 are independently Ci-Cig alkyl or (Co-C 4 alkyl)(C 6 -Cio aryl)R 7 ;
  • R 3 is Ci-Ci 8 alkyl or R 3 and R 4 together with the atoms to which they are attached form a 4-12 heterocyclic ring;
  • R 4 and R 8 are independently selected from the group consisting of hydrogen, Ci-Cie alkyl and (C 0 -C 4 alkyl)(C 6 -C 10 aryl)R 7 ;
  • R 5 is NH 2 or OH
  • R 7 is selected from the group consisting of hydrogen, Ci-Cig alkyl, C 2 -Cig alkenyl, (C 0 -C 4 alkyl)CONH 2> (C 0 -C 4 alkyl)COOH, (C 0 -C 4 alkyl)NH 2 , (C 0 -C 4 alkyl)OH, and halo.
  • A-B comprises the structure of formula I linked via an amide bond to an amine substituent on an aryl of Q of the complex A-B-Q, wherein Ri is selected from the group consisting of hydrogen, Q-Cig alkyl and aryl, or
  • Ri and R 2 are linked through -(CH 2 ) P -, wherein p is 2-9;
  • R 3 is Ci-Cig alkyl or R 3 and R 4 together with the atoms to which they are attached form a 4-6 heterocyclic ring;
  • R 4 and R 8 are independently selected from the group consisting of hydrogen, Ci-Qg alkyl and aryl;
  • R 5 is an amine or N-substituted amine.
  • the dipeptide element comprises the structure of formula I linked via an amide bond to an amine substituent on an aryl of Q of the complex A-B-Q, wherein
  • Ri is selected from the group consisting of hydrogen, Q-Qg alkyl, (C Cig alkyl)OH, (C1-C4 alkyl)NH 2 , and (C 0 -C 4 alkyl)(C 6 -C 10 aryl)R 7 ;
  • R 3 is Ci-Cig alkyl or R 3 and R 4 together with the atoms to which they are attached form a 4-6 heterocyclic ring;
  • R 4 and Rg are independently selected from the group consisting of hydrogen, C1-C18 alkyl and (C 0 -C 4 alkyl)(C 6 -C 10 aryl)R 7 ;
  • R 5 is NHR 6 ;
  • R 6 is H, Q-Cg alkyl, or R 6 and Ri together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;
  • R 7 is selected from the group consisting of hydrogen, Q-Qg alkyl, C 2 -Cig alkenyl, (C 0 -C 4 alkyl)CONH 2> (C 0 -C 4 alkyl)COOH, (C 0 -C 4 alkyl)NH 2 , (C 0 -C 4 alkyl)OH, and halo.
  • the dipeptide element (A-B) comprises the structure of formula I linked via an amide bond to an amine substituent on an aryl of Q of the complex A-B-Q, wherein
  • Ri and R 2 are independently selected from the group consisting of hydrogen, C Cg alkyl and aryl;
  • R 3 is Ci-Cig alkyl or R 3 and R 4 together with the atoms to which they are attached form a 4-6 heterocyclic ring;
  • R 4 and R 8 are each hydrogen
  • R5 is selected from the group consisting of amine, N-substituted amine and hydroxyl.
  • the dipeptide element is linked via an amide bond to an amine substituent on an aryl and comprises the structure:
  • R and R 2 are independently selected from the group consisting of hydrogen, Ci-C 8 alkyl, (C1-C4 alkyl)COOH, and (C 0 -C 4 alkyl)(C 6 -Ci 0 aryl)R 7> or Rj and R 5 together with the atoms to which they are attached form a 4-11 heterocyclic ring;
  • R3 is Ci-C 18 alkyl or R 3 and R 4 together with the atoms to which they are attached form a 4-6 heterocyclic ring;
  • R 4 and R 8 are each hydrogen
  • R 5 is NHR 6 or OH
  • R 6 is H or Ci-C 8 alkyl, or R 6 and R together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;
  • R 7 is selected from the group consisting of hydrogen, Ci-C 18 alkyl, C 2 -C 18 alkenyl, (C 0 -C 4 alkyl)CONH 2, (C 0 -C 4 alkyl)COOH, (C 0 -C 4 alkyl)NH 2 , (C 0 -C 4 alkyl)OH, and halo.
  • Q is a medicinal agent and in one embodiment Q is a compound selected from the group consisting of thyroxine T4 (3,5,3',5'-tetraiodothyronine), 3,5,3'-triiodo L-thyronine and 3,3',5'-triiodo L- thyronine.
  • the dipeptide/drug complex comprises the structure of
  • R ⁇ R 2> R ⁇ and R 8 are independently selected from the group consisting of H, C1-C18 alkyl, C 2 -Ci 8 alkenyl, (Ci-Ci 8 alkyl)OH, (Ci-Ci 8 alkyl)SH, (C 2 -C 3 alkyl)SCH 3 , (C1-C4 alkyl)CONH 2 , (C1-C4 alkyl)COOH, (C1-C4 alkyl)NH 2 , (C1-C4
  • R 3 is selected from the group consisting of Ci-C 18 alkyl, (Ci-C 18 alkyl)OH, (C 1 -C18 alkyl)NH 2 , (Ci-Cie alkyl)SH, (C 0 -C 4 alkyl)(C 3 -C 6 )cycloalkyl, (C 0 -C 4 alkyl)(C 2 -C 5 heterocyclic), (C 0 -C 4 alkyl)(C 6 -Ci 0 aryl)R 7 , and (C 1 -C4 alkyl)(C 3 -C 9 heteroaryl) or R 4 and R 3 together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;
  • R 5 is NHR 6 or OH
  • R 6 is H, Q-Cg alkyl or R 6 and R 2 together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;
  • R 7 is selected from the group consisting of H and OH
  • R 15 and R 16 are independently selected from hydrogen and iodine.
  • the dipeptide/drug complex comprises the structure of Formul
  • R ⁇ R 2j R4 and R 8 are independently selected from the group consisting of H, C 1 -C18 alkyl, C 2 -Ci 8 alkenyl, (Ci-Ci 8 alkyl)OH, (Ci-Ci 8 alkyl)SH, (C 2 -C 3 alkyl)SCH 3 , (C1-C4 alkyl)CONH 2 , (C1-C4 alkyl)COOH, (C1-C4 alkyl)NH 2 , (C1-C4
  • R 3 is selected from the group consisting of Ci-C 18 alkyl, (Ci-C 18 alkyl)OH, (Ci-Cie alkyl)NH 2 , (Ci-C 18 alkyl)SH, (C 0 -C 4 alkyl)(C 3 -C 6 )cycloalkyl, (C 0 -C 4 alkyl)(C 2 -C 5 heterocyclic), (C 0 -C 4 alkyl)(C 6 -C 10 aryl)R 7 , and (C1-C4 alkyl)(C 3 -C 9 heteroaryl) or R 4 and R 3 together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;
  • R 5 is NHR 6 or OH
  • R 6 is H, Q-C8 alkyl or R 6 and Ri together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;
  • R 7 is selected from the group consisting of hydrogen, Ci-Cig alkyl, C 2 -Cig alkenyl, (C 0 -C 4 alkyl)CONH 2, (C 0 -C 4 alkyl)COOH, (C 0 -C 4 alkyl)NH 2 , (C 0 -C 4 alkyl)OH, and halo; and
  • Ri 5 and R 16 are independently selected from hydrogen and iodine.
  • Ri is selected from the group consisting of H and Ci-Cg alkyl
  • R 2 and R 4 are independently selected from the group consisting of H, Ci-C 8 alkyl, C 2 -C 8 alkenyl, (Ci-C 4 alkyl)OH, (Ci-C 4 alkyl)SH, (C 2 -C 3 alkyl)SCH 3 , (Ci-C 4 alkyl)CONH 2 , (C1-C4 alkyl)COOH, (C1-C4 alkyl)NH 2 , (C1-C4
  • R 3 is selected from the group consisting of Ci-Cg alkyl, (C 1 -C 4 alkyl)OH, (C1-C4 alkyl)NH 2> (C1-C4 alkyl)SH, (C 3 -C 6 )cycloalkyl or R 4 and R 3 together with the atoms to which they are attached form a 5 or 6 member heterocyclic ring;
  • R 5 is NHR 6 or OH
  • R is H, or R and R 2 together with the atoms to which they are attached form a 5 or 6 member heterocyclic ring;
  • R 7 is selected from the group consisting of H and OH; and R8 is H, with the proviso that when R 4 and R 3 together with the atoms to which they are attached form a 5 or 6 member heterocyclic ring, at least one of Ri and R 2 are not H, and in one embodiment both Ri and R 2 are other than H.
  • Ri is H or Ci-C 8 alkyl
  • R 2 and R 4 are independently selected from the group consisting of H, Ci-Cg alkyl, C 2 -C 8 alkenyl, (C1-C4 alkyl)OH, (C1-C4 alkyl)SH, (C 2 -C 3 alkyl)SCH 3 , (C1-C4 alkyl)CONH 2 , (C 1 -C4 alkyl)COOH, (C 1 -C4 alkyl)NH 2 , (C 1 -C4 alkyl)NHC(NH 2 +) NH 2 , (C 0 -C4 alkyl)(C 3 -C 6 cycloalkyl), (C 0 -C 4 alkyl)(C 2 -C 5 heterocyclic), (C 0 -C 4 alkylXCe-Cio aryl)R7, and CH 2 (C 3 -C9 heteroaryl), or R and R 2 together with the atoms to which they are attached form a C 3 -C 12
  • R 3 is C 1 -C18 alkyl; (C 1 -C4 alkyl)OH, (C 1 -C4 alkyl)NH 2> (C 1 -C4 alkyl)SH, (C 3 -
  • R 5 is NHR 6 or OH
  • R 6 is H or R 6 and R 2 together with the atoms to which they are attached form a 5 or 6 member heterocyclic ring;
  • R 7 is selected from the group consisting of hydrogen, Q-C ⁇ alkyl, C 2 -Ci 8 alkenyl, (C 0 -C 4 alkyl)CONH 2, (C 0 -C 4 alkyl)COOH, (C 0 -C 4 alkyl)NH 2 , (C 0 -C 4 alkyl)OH, and halo; and
  • Rg is H, with the proviso that when R 4 and R together with the atoms to which they are attached form a 5 or 6 member heterocyclic ring, at least one of R and R 2 are not H, and in one embodiment both R and R 2 are other than H.
  • any of the complexes disclosed herein can be further modified to improve the peptide's solubility in aqueous solutions at physiological pH, while enhancing the effective duration of the peptide by preventing renal clearance of the peptide.
  • the peptide prodrugs are further modified to comprise a covalently linked hydrophilic moiety.
  • the hydrophilic moiety is a plasma protein, polyethylene glycol chain or the Fc portion of an immunoglobin. Therefore, in one embodiment the presently disclosed complexes are further modified to comprise one or more hydrophilic groups covalently linked to the side chain of the dipeptide element A-B, or optional to other amino acid side chains when the medicinal agent is a bioactive peptide.
  • the dipeptide/drug complexes are modified to comprise an acyl group or alkyl group.
  • Acylation or alkylation can increase the half-life of the drug in circulation.
  • Acylation or alkylation can advantageously delay the onset of action and/or extend the duration of action at the drugs target receptor and/or improve resistance to proteases such as DPP-IV.
  • Acylation may also enhance solubility of the dipeptide/drug complex at neutral pH.
  • an amino acid of the dipeptide element A-B is acylated.
  • the acyl group can be covalently linked directly to the medicinal agent, or indirectly to the medicinal agent via a spacer, wherein the spacer is positioned between the medicinal agent and the acyl group.
  • the medicinal agent comprises an amino acid
  • the medicinal agent is acylated through the side chain amine, hydroxyl, or thiol of an amino acid of the medicinal agent. Suitable methods of peptide acylation via amines, hydroxyls, and thiols are known in the art. See, for example, Miller, Biochem Biophys Res Commun 218: 377-382 (1996);
  • the acyl group of the acylated medicinal agent can be of any size, e.g., any length carbon chain, and can be linear or branched.
  • the acyl group is a C4 to C28 fatty acid.
  • the acyl group can be any of a C4 fatty acid, C6 fatty acid, C8 fatty acid, CIO fatty acid, C12 fatty acid, C14 fatty acid, C16 fatty acid, C18 fatty acid, C20 fatty acid, C22 fatty acid, C24 fatty acid, C26 fatty acid, or a C28 fatty acid.
  • the acyl group is a C8 to C20 fatty acid, e.g., a C14 fatty acid or a C16 fatty acid.
  • the acyl group is a fatty acid or bile acid, or salt thereof, e.g. a C4 to C30 fatty acid, a C8 to C24 fatty acid, cholic acid, a C4 to C30 alkyl, a C8 to C24 alkyl, or an alkyl comprising a steroid moiety of a bile acid.
  • the amino acid at the position of the dipeptide element A- B where the hydrophilic moiety is to be linked is selected to allow for ease in attaching the hydrophilic moiety.
  • the dipeptide element may comprise a lysine or cysteine residue to allow for the covalent attachment of a polyethylene glycol chain.
  • the dipeptide/drug complex has a single cysteine residue, present in the dipeptide element A-B, wherein the side chain of the cysteine residue is further modified with a thiol reactive reagent, including for example, maleimido, vinyl sulfone, 2-pyridylthio, haloalkyl, and haloacyl.
  • thiol reactive reagents may contain carboxy, keto, hydroxyl, and ether groups as well as other hydrophilic moieties such as polyethylene glycol units.
  • the complex has a single lysine residue, present in the dipeptide element A-B, and the side chain of the substituting lysine residue is further modified using amine reactive reagents such as active esters (succinimido, anhydride, etc) of carboxylic acids or aldehydes of hydrophilic moieties such as polyethylene glycol.
  • amine reactive reagents such as active esters (succinimido, anhydride, etc) of carboxylic acids or aldehydes of hydrophilic moieties such as polyethylene glycol.
  • the dipeptide/drug complex comprises a polyethylene glycol chain
  • the polyethylene glycol chain may be in the form of a straight chain or it may be branched.
  • the polyethylene glycol chain has an average molecular weight selected from the range of about 20,000 to about 60,000 Daltons.
  • Multiple polyethylene glycol chains can be linked to the prodrugs to provide a prodrug with optimal solubility and blood clearance properties.
  • the dipeptide/drug complex is linked to a single polyethylene glycol chain that has an average molecular weight selected from the range of about 20,000 to about 60,000 Daltons.
  • the dipeptide/drug complex is linked to a two polyethylene glycol chains wherein the combined average molecular weight of the two chains is selected from the range of about 40,000 to about 80,000 Daltons.
  • a single polyethylene glycol chain having an average molecular weight of 20,000 or 60,000 Daltons is linked to the dipeptide/drug complex.
  • a single polyethylene glycol chain is linked to the dipeptide/drug complex and has an average molecular weight selected from the range of about 40,000 to about 50,000 Daltons.
  • two polyethylene glycol chains are linked to the dipeptide/drug complex wherein the first and second polyethylene glycol chains each have an average molecular weight of 20,000 Daltons.
  • two polyethylene glycol chains are linked to the dipeptide/drug complex wherein the first and second polyethylene glycol chains each have an average molecular weight of 40,000 Daltons.
  • a medicinal prodrug analog wherein a plasma protein has been covalently linked to an amino acid side chain of the dipeptide element, or optionally to another amino acid side chain when the medicinal agent is a bioactive peptide, to improve the solubility, stability and/or pharmacokinetics of the prodrug.
  • a plasma protein has been covalently linked to an amino acid side chain of the dipeptide element, or optionally to another amino acid side chain when the medicinal agent is a bioactive peptide, to improve the solubility, stability and/or pharmacokinetics of the prodrug.
  • one or more serum albumins can be covalently bound, or non-covalently bound via a high affinity association (e.g. via a C16-C18 acylated amino acid side chain) to the dipeptide/medicinal agent complex.
  • a dipeptide/medicinal agent complex wherein a linear amino acid sequence representing the Fc portion of an immunoglobin molecule has been covalently linked to an amino acid side chain of the dipeptide element, or optionally to another amino acid side chain when the medicinal agent is a bioactive peptide, to improve the solubility, stability and/or
  • the Fc portion is typically one isolated from IgG, but the Fc peptide fragment from any immunoglobin should function equivalently.
  • the present disclosure also encompasses other conjugates in which prodrugs of the invention are linked, optionally via covalent bonding and optionally via a linker, to a conjugate moiety.
  • Linkage can be accomplished by covalent chemical bonds, physical forces such electrostatic, hydrogen, ionic, van der Waals, or hydrophobic or hydrophilic interactions.
  • a variety of non-covalent coupling systems may be used, including biotin-avidin, ligand/receptor, enzyme/substrate, nucleic acid/nucleic acid binding protein, lipid/lipid binding protein, cellular adhesion molecule partners; or any binding partners or fragments thereof which have affinity for each other.
  • conjugates include but are not limited to a heterologous peptide or polypeptide (including for example, a plasma protein), a targeting agent, an immunoglobulin or portion thereof (e.g. variable region, CDR, or Fc region), a diagnostic label such as a radioisotope, fluorophore or enzymatic label, a polymer including water soluble polymers, or other therapeutic or diagnostic agents.
  • a conjugate is provided comprising a prodrug of the present invention and a plasma protein, wherein the plasma protein is selected form the group consisting of albumin, transferin and fibrinogen.
  • the plasma protein moiety of the conjugate is albumin or transferin.
  • the linker may comprises a chain of atoms from 1 to about 60, or 1 to 30 atoms or longer, 2 to 5 atoms, 2 to 10 atoms, 5 to 10 atoms, or 10 to 20 atoms long.
  • the chain atoms are all carbon atoms.
  • the chain atoms in the backbone of the linker are selected from the group consisting of C, O, N, and S. Chain atoms and linkers may be selected according to their expected solubility (hydrophilicity) so as to provide a more soluble conjugate.
  • the linker provides a functional group that is subject to cleavage by an enzyme or other catalyst or hydrolytic conditions found in the target tissue or organ or cell.
  • the length of the linker is long enough to reduce the potential for steric hindrance. If the linker is a covalent bond or a peptidyl bond and the conjugate is a polypeptide, the entire conjugate can be a fusion protein.
  • Such peptidyl linkers may be any length. Exemplary linkers are from about 1 to 50 amino acids in length, 5 to 50, 3 to 5, 5 to 10, 5 to 15, or 10 to 30 amino acids in length.
  • Such fusion proteins may alternatively be produced by recombinant genetic engineering methods known to one of ordinary skill in the art.
  • the dipeptide prodrug element can be further modified to comprise a hydrophilic moiety.
  • the hydrophilic moiety is a polyethylene glycol chain.
  • a polyethylene glycol chain of 40k or higher is covalently bound to the side chain of the A or B amino acid of the dipeptide prodrug element.
  • the dipeptide prodrug element is additionally or alternatively acylated or alkylated with a fatty acid or bile acid, or salt thereof, e.g.
  • the 'A' amino acid of the dipeptide prodrug element can include, for example, d-lysine covalently bound to an acyl or alkyl group through its side chain amino group, or d-cysteine covalently bound to a PEG molecule through its side chain sulfhydryl group.
  • the dipeptide prodrug element can be directly bound to the hydrophilic moiety, acyl group, or alkyl group, or bound to the hydrophilic moiety, acyl group, or alkyl group through a spacer, as described herein.
  • the dipeptide prodrug element can be linked to a depot protein such as dextran or a large PEG molecule (greater or equal to 80,000 daltons) that serves to sequester the prodrug at an injection site until cleavage of the dipeptide releases the active bioactive peptide
  • the rate of cleavage of the dipeptide prodrug element A-B from the bioactive peptide, and thus activation of the prodrug depends on the structure (including N-alkylation, number of substituents, length or bulkiness), and stereochemistry of the amino acids of the dipeptide prodrug element.
  • the rate of cleavage of the dipeptide prodrug element A-B from the bioactive peptide also depends on the steric hindrance, nucleophilicity, and stability of the leaving group of Q during diketopiperazine formation.
  • peptide sequences disclosed in Int'l Application No. PCT/US2009/68745, filed December 18, 2009 or its sequence listing and sub-categories of (1) dipeptide prodrug elements, (2) A amino acids, and/or (3) B amino acids disclosed in Int'l Application No. PCT/US2009/68745, filed December 18, 2009, to the extent they fall completely within and/or overlap with a portion of any of the sub-categories described herein, and only to the extent necessary to confer novelty on claimed subject matter.
  • the half-life of the prodrug is dependent on the presence and length of the N-alkyl substituent on the B amino acid.
  • a prodrug that has a shorter N-alkyl substituent on the B amino acid e.g. Gly(N-methyl)
  • the half-life of the prodrug is dependent on the presence or absence of an alkyl side chain, and the degree of substitution at the beta position of the alkyl side chain, of the B amino acid of the dipeptide prodrug element.
  • a prodrug that has an N-alkylated B amino acid that is disubstituted at the beta position e.g., N-alkylated isoleucine
  • will undergo slower cleavage of A-B, and have a longer half-life, than a prodrug that has an N-alkylated B amino acid that is monosubstituted at the beta position e.g., N-alkylated leucine.
  • a prodrug that has an N-alkylated B amino acid that is monosubstituted at the beta position will undergo slower cleavage of A-B, and have a longer half-life, than a prodrug that has an N-alkylated B amino acid that is unsubstituted at the beta position (e.g., N-alkylated alanine).
  • a prodrug with an N-alkylated B amino acid that has an unsubstituted beta position (e.g., N-alkylated alanine) will undergo slower cleavage of A-B, and have a longer half-life, than a prodrug that has glycine or N-alkylated glycine as the B amino acid.
  • the half-life of the prodrug is dependent on the bulkiness of the side chain of the B amino acid.
  • a prodrug that has a bulkier side chain on the B amino acid e.g., N-alkylated phenylalanine
  • Cleavage rates of dipeptides can be further differentiated by the amine of the drug (e.g., insulin) to which they are attached.
  • dipeptide linked to an N-terminal amine will cleave at a faster rate relative to when the dipeptide is linked to the side chain amine of a lysine residue.
  • composition of the B amino acid of the dipeptide prodrug element can be classified into the below sub-categories IA, IB, and IC.
  • the dipeptide prodrug elements in sub-category IA undergo cleavage the fastest and the dipeptide prodrug elements in sub-category IC undergo cleavage the slowest.
  • Sub-Category IA Amino Acid B of the Dipeptide Prodrug Element is N- Alkylated Glycine
  • the prodrug comprises the structure:
  • Q is a bioactive peptide (e.g., an insulin peptide);
  • A-B comprises the structure:
  • Ri and R 2 are independently selected from the group consisting of H, C C ⁇ alkyl, C 2 -C 18 alkenyl, (Ci-Cie alkyl)OH, (Ci-Cie alkyl)SH, (C 2 -C 3 alkyl)SCH 3 , (C C alkyl)CONH 2 , (C 1 -C4 alkyl)COOH, (C 1 -C4 alkyl)NH 2 , (C 1 -C4 alkyl)NHC(NH 2 + )NH 2 (C0-C4 alkyl)(C 3 -C 6 cycloalkyl), (C 0 -C 4 alkyl)(C 2 -C 5 heterocyclic), (C 0 -C 4 alkyl)(C 6 - C10 aryl)R 7 , (C1-C4 alkyl)(C 3 -C 9 heteroaryl), and C1-C12 alkyl(Wi)Ci-Ci 2 alkyl, where
  • R 3 is Ci-Cie alkyl
  • R 4 and R 8 are each H
  • R 5 is NHR 6 ;
  • R is H or CrC 4 alkyl, or R 5 and R 2 together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;
  • R 7 is selected from the group consisting of H and OH.
  • the B amino acid is selected from the group consisting of glycine(N-methyl), glycine(N-ethyl), glycine(N-propyl), glycine(N-butyl), glycine (N-pentyl), glycine(N-hexyl), glycine(N-heptyl), and glycine(N-octyl).
  • the B amino acid can be glycine(N-methyl) or glycine(N-hexyl).
  • R 3 is Ci-C 4 alkyl. In some embodiments when one of Ri or R 2 is other than hydrogen, R 3 is CrC 4 alkyl.
  • Sub-Category IB Amino Acid B of the Dipeptide Prodrug Element is
  • the prodrug comprises the structure:
  • Q is a bioactive peptide (e.g., an insulin peptide);
  • A-B comprises the structure:
  • Ri and R 2 are independently selected from the group consisting of H, Ci-Cig alkyl, C 2 -Ci 8 alkenyl, (Ci-C 18 alkyl)OH, (Ci-C 18 alkyl)SH, (C 2 -C 3 alkyl)SCH 3 , (C C 4 alkyl)CONH 2 , (Ci-C 4 alkyl)COOH, (Ci-C 4 alkyl)NH 2 , (Ci-C 4
  • R 4 is selected from the group consisting of CH 3 , CH 2 (C 1 -C 1 o alkyl), CH 2 (C 2 - C10 alkenyl), CH 2 (C 0 -C 10 alkyl)OH, CH 2 (C 0 -C 10 alkyl)SH, CH 2 (C 0 -C 3 alkyl)SCH 3 , CH 2 (C 0 -C 3 alkyl)CONH 2 , CH 2 (C 0 -C 3 alkyl)COOH, CH 2 (C 0 -C 3 alkyl)NH 2 , CH 2 (C 0 - C 3 alkyl)NHC(NH 2 + )NH 2 , CH 2 (C 0 -C 3 alkyl)(C 3 -C 6 cycloalkyl), CH 2 (C 0 -C 3 alkyl)(C 2 -C 5 heterocyclic), CH 2 (C 0 -C 3 alkyl)(C 6 -Ci 0 aryl)R
  • R 8 is H
  • R 5 is NHR 6 , or R 5 and R 2 together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;
  • R 6 is H or Ci-C 4 alkyl
  • R 7 is selected from the group consisting of H and OH.
  • R 4 is selected from the group consisting of CH 3>
  • Nonlimiting examples of the B amino acid in these embodiments include alanineiN-Ci-Cioalkyl), leucineiN-CrCioalkyl), methionineiN-CrCioalkyl), asparagineiN-Ci-Cioalkyl), glutamic acidiN-Ci-Cioalkyl), aspartic acid(N-Ci- C 10 alkyl), glutamineiN-CrCioalkyl), histidineiN-CrCioalkyl), lysineiN-Cr Cioalkyl), arginineiN-CrCioalkyl), serineiN-CrCioalkyl), and cysteineiN-Cr Cioalkyl).
  • the B amino acid is selected from the group consisting of alanineiN-Ci-Cealkyl), leucineiN-Q-Cealkyl), methionineiN-CrCealkyl), asparagineiN-Ci-Cealkyl), glutamic acid(N-Ci-C 6 alkyl), aspartic acid(N-Cr C 6 alkyl), glutamineCN-CrCealkyl), histidineiN-CrCealkyl), lysineiN-CrCealkyl), arginineiN-Ci-Cealkyl), serineiN-C Cealkyl), and cysteineiN-Ci-Cealkyl).
  • the B amino acid can include alanine(N-methyl), leucine(N- methyl), methionine(N-methyl), asparagine(N-methyl), glutamic acid(N-methyl), aspartic acid(N-methyl), glutamine(N-methyl), histidine(N-methyl), lysine(N- methyl), arginine(N-methyl), serine(N-methyl), and cysteine(N-methyl).
  • R 4 is selected from the group consisting of CH 2 (Co-C3 alkyl)(C 3 -C 6 cycloalkyl), CH 2 (C 0 -C 3 alkyl)(C 2 -C 5 heterocyclic), CH 2 (C 0 -C 3 alkyl)(C 6 -Cio aryl)R 7 , CH 2 (Ci-C 3 alkyl)(C 3 -C 9 heteroaryl), and CH 2 (C 0 -C 12 alkylXW C Cn alkyl, wherein Wi is a heteroatom selected from the group consisting of N, S and O, and wherein R 7 is selected from the group consisting of H and OH.
  • Nonlimiting examples of the B amino acid in these embodiments include phenylalanineiN-Ci-Cioalkyl), tyrosineiN-CrCioalkyl), and tryptophan(N-Ci- Cioalkyl).
  • the B amino acid is selected from the group consisting of phenylalanineiN-Ci-Cealkyl), tyrosineiN-Ci-Cealkyl), and
  • the B amino acid can include
  • the B amino acid is proline.
  • proline is excluded from Sub-Category IB.
  • Sub-Category IC Amino Acid B of the Dipeptide Prodrug Element
  • the prodrug comprises the structure:
  • A-B-Q wherein Q is a bioactive peptide (e.g., an insulin peptide); wherein A-B comprises the structure:
  • Ri and R 2 are independently selected from the group consisting of H, Ci-Ci 8 alkyl, C 2 -Ci8 alkenyl, (Ci-Ci 8 alkyl)OH, (Ci-Ci 8 alkyl)SH, (C 2 -C 3 alkyl)SCH 3 , (C C 4 alkyl)CONH 2 , (C1-C4 alkyl)COOH, (C1-C4 alkyl)NH 2 , (C1-C4
  • R 4 is independently selected from the group consisting of CH(Ci-C8 alkyl) 2 , CH (C 2 -C 8 alkenyl) 2 , CH(Ci-C 8 alkyl)(OH), CH(Ci-C 8 alk l)((Ci-C 8 alkyl)SH), CH(d-C 3 alkylXCQ-Cg alkyl)(NH 2) );
  • R 8 is H;
  • R 5 is NHR 6j or R 5 and R 2 together with the atoms to which they are attached form a 4, 5 or 6 member heterocyclic ring;
  • R 6 is H or Ci-C 4 alkyl
  • R 7 is selected from the group consisting of H and OH.
  • R 4 is CH(Ci-C 8 alkyl) 2 or CH(Ci-C 8 alkyl)OH.
  • Nonlimiting examples of the B amino acid include isoleucine(N-Ci-Cioalkyl), valineiN-Ci-Cioalkyl), and threonineiN-Ci-Cioalkyl).
  • the B amino acid is selected from the group consisting of isoleucine(N-Ci-C 6 alkyl), valine(N-Ci-C 6 alkyl), and threonine(N-Ci-C 6 alkyl).
  • the B amino acid can include isoleucine(N-methyl), valine(N-methyl), and threonine(N-methyl).
  • the half-life of the prodrug is dependent on the number of substituents at the alpha position of the A amino acid.
  • a prodrug comprising an A amino acid that is an a-monosubstituted amino acid e.g., Ala
  • a prodrug comprising an A amino acid that is an ⁇ , ⁇ -disubstituted amino acid e.g., Aib
  • the half-life of the prodrug is dependent on the degree of alkylation on the alpha amino group of the A amino acid. Generally, the greater the degree of alkylation, the slower the rate of cleavage and the longer the half-life of the prodrug. For example, a dipeptide prodrug element having N-alkylated Ala will cleave at a slower rate, and have a longer half-life, than Ala.
  • composition of the A amino acid of the dipeptide prodrug element can be classified into the below sub-categories IIA and IIB.
  • dipeptide prodrug elements in sub-category IIA cleave faster than dipeptide prodrug elements in subcategory IIB.
  • Sub-Category IIA Amino Acid A of the Dipeptide Prodrug Element is Disubstituted at the Alpha Position
  • the A amino acid of the dipeptide prodrug element is disubstituted at the alpha position.
  • Ri and R 2 of the structures described in sub-categories IA, IB, and IC are independently selected from the group consisting of Ci-Cio alkyl, C 2 -Ci 0 alkenyl, (Ci-Cio alkyl)OH, (Ci-Cio alkyl)SH, (C 2 - C 3 alkyl)SCH 3 , (Ci-C 4 alkyl)CONH 2 , (Ci-C 4 alkyl)COOH, (Ci-C 4 alkyl)NH 2 , (Ci-C 4 alkyl)NHC(NH 2 + )NH 2 , (C 0 -C 4 alkyl)(C 3 -C 6 cycloalkyl), (C 0 -C 4 alkyl)(C 2 -C 5 heterocyclic), (C 0 -C 4 alkyl
  • the A amino acid can include aminoisobutyric acid (Aib).
  • Sub-Category IIB Amino Acid A of the Dipeptide Prodrug Element is Unsubstituted or Monosubstituted at the Alpha Position
  • the A amino acid of the dipeptide prodrug element is unsubstituted or monosubstituted at the alpha position.
  • Ri of the structures described in sub-categories IA, IB, and IC is H
  • R 2 of the structures described in sub-categories IA, IB, and IC is selected from the group consisting of H, Ci-Cio alkyl, C 2 -C 10 alkenyl, (Ci-Cio alkyl)OH, (Ci-Cio alkyl)SH, (C 2 -C 3 alkyl)SCH 3 , (Ci-C 4 alkyl)CONH 2 , (C C 4 alkyl)COOH, (C C 4 alkyl)NH 2 , (C C 4 alkyl)NHC(NH 2 + )NH 2 , (C 0 -C 4 alkyl)(C 3 -C 6 cycloalkyl), (C 0 -C 4 alkyl)
  • the A amino acid of the dipeptide prodrug element has 'd' stereochemistry.
  • Nonlimiting examples of the A amino acid in these embodiments include lysine, cysteine, and alanine.
  • d- lysine, d-cysteine, and d- alanine may enhance half-life through reducing proteolytic degradation of the prodrug peptide.
  • the A amino acid is N-alkylated with a group that has 1 to 4 carbon atoms such as Ala(N-Ci-C 4 alkyl), Lys(N-Ci-C 4 alkyl), and Cys(N-Ci- C 4 alkyl).
  • the A amino acid can be Ala(N-methyl), Lys(N-methyl), and Cys(N-methyl). N-alkylation of the A amino acid decreases the rate of cleavage of the dipeptide prodrug element from Q and provides a longer half-life.
  • the half-life of the prodrug depends on the steric hindrance, nucleophilicity, and stability of the leaving group on Q during
  • the type of leaving group on Q can be determined by the type of the linkage between A-B and an amino group of Q, as described in sub-categories IIIA and IIIB below. Generally, dipeptide prodrug elements in sub-category IIIB cleave faster from Q and have a shorter half-life than dipeptide prodrug elements in subcategory IIIA.
  • Sub-Category IIIA A-B Linked to an Aliphatic Amino Group of Q
  • A-B is linked to Q through an amide bond between A- B and an aliphatic amino group of Q to result in a prodrug with a chemical cleavage half-life (t 1/2 ) of A-B from Q of at least about 1 hour to about 1 week in PBS, under physiological conditions, as previously described herein.
  • A-B is linked to Q through an amide bond between A- B and the alpha amino group of the N-terminal amino acid of Q.
  • a dipeptide prodrug element having a B amino acid from any of sub-categories IA, IB, and IC and an A amino acid from any of sub-categories IIA and IIB can be linked to the N-terminal amino acid of Q to result in a prodrug with a chemical cleavage half- life (ti /2 ) of A-B from Q of at least about 1 hour to about 1 week in PBS, under physiological conditions.
  • A-B is linked to Q through an amide bond between A-
  • a dipeptide prodrug element having a B amino acid from any of sub-categories IA, IB, and IC and an A amino acid from any of sub-categories IIA and IIB can be linked to an aliphatic amino group of a side chain of an amino acid of Q to result in a prodrug with a chemical cleavage half-life (ti /2 ) of A-B from Q of at least about 1 hour to about 1 week in PBS, under physiological conditions.
  • A-B when A-B is linked to Q through an amide bond between A-B and an aliphatic amino group of Q, either A should be an ⁇ , ⁇ - disubstituted amino acid (Sub-category IIA) or B should be N-alkylated (any of Sub- categories IA, IB or IC) , or both.
  • A is an a-monosubstituted amino acid (e.g., Ala)
  • B is not N-alkylated, and A-B is attached to Q through an aliphatic amino group of Q, then there will not be significant cleavage of A-B.
  • the N-alkyl substitutent of the B amino acid has a length of at least five carbon atoms (for example, N-Cs-Csalkyl).
  • the B amino acid is not proline.
  • Sub-Category IIIB A-B Linked to an Aromatic Amino Group of Q
  • A-B is linked to Q through an amide bond between A- B and an aromatic amino group of a side chain of an amino acid of Q to result in a prodrug with a chemical cleavage half-life (t ⁇ ) of A-B from Q of at least about 1 hour to about 1 week in PBS, under physiological conditions, as previously described herein.
  • a dipeptide prodrug element having a B amino acid from any of sub-categories IA, IB, and IC and an A amino acid from any of sub-categories IIA and IIB can be linked to an aromatic amino group of a side chain of an amino acid of Q to result in a prodrug with a chemical cleavage half-life (t ⁇ ) of A-B from Q of at least about 1 hour to about 1 week in PBS, under physiological conditions.
  • t ⁇ chemical cleavage half-life
  • Any of the B amino acids defined by Category I can be combined with any of the A amino acids defined by Category II to form a dipeptide prodrug element.
  • This dipeptide prodrug element can be linked to any of the positions described in Category III.
  • the half life of the prodrug can be tuned through the selection of:
  • the dipeptide prodrug elements described above can be further modified to comprise a hydrophilic moiety, an acyl group, or an alkyl group, as previously described herein.
  • the dipeptide prodrug element includes lysine that is conjugated to an acyl group or an alkyl group through its side chain amino group.
  • the dipeptide prodrug element includes cysteine that is conjugated to a hydrophilic moiety (e.g., 40 kD PEG) through the side chain sulfhydryl group.
  • the hydrophilic moiety, acyl group, or alkyl group can be conjugated directly to the dipeptide prodrug element or through a spacer.
  • the hydrophilic group, the alkyl group and/or the acyl group are conjugated to the A amino acid of the dipeptide prodrug element.
  • the following dipeptide prodrug elements include an acyl group: dLys-Gly(N-Hexyl), dLys-Gly(N-Methyl), and dLys-Phe(N-Methyl).
  • the following dipeptide prodrug elements include an alkyl group: dLys-Gly(N-Hexyl), dLys-Gly(N-Methyl), and dLys-Phe(N-Methyl).
  • the dipeptide prodrug element of the invention can include combinations of any of the B amino acids from Category I with any of the A amino acids from Category II.
  • Nonlimiting examples of amino acids suitable for the A amino acid and for the B amino acid of the dipeptide prodrug element are listed in the below Table.
  • the dipeptide prodrug element includes the
  • combinations of the A amino acid and the B amino acid of the dipeptide prodrug element can include: Al-Bl; A1-B2; A1-B3; A1-B4; A1-B5; A1-B6; A1-B7; A1-B8; A1-B9; A1-B10; Al-Bl l ; A1-B12; A1-B13; A1-B14; A1-B15; A1-B16; A1-B17; A1-B18; A1-B19; A1-B20; A1-B21; A1-B22; A1-B23; A1-B24; A1-B25; A1-B26; A1-B27; A1-B28; A1-B29; A1-B30; A1-B31; A1-B32; A1-B33; A1-B34; A1-B
  • the dipeptide prodrug element includes the
  • combinations of the A amino acid and the B amino acid of the dipeptide prodrug element can include: Al-Bl; A1-B2; A1-B3; A1-B4; A1-B5; A1-B6; A1-B7; A1-B8; A1-B9; A1-B10; Al-Bl l ; A1-B12; A1-B13; A1-B14; A1-B 15; A1-B16; A1-B17;

Abstract

L'invention porte sur un élément dipeptide pouvant subir une séparation automatique par voie non enzymatique, ledit dipeptide pouvant être lié à des agents médicinaux connus par une liaison amide. Le dipeptide sera spontanément séparé de l'agent médicinal dans certaines conditions physiologiques par une réaction entraînée par une instabilité chimique. Par conséquent, l'élément dipeptide fournit un moyen de liaison de divers composés à des agents médicinaux connus, les composés étant par la suite libérés de l'agent médicinal après un temps d'exposition prédéfini à certaines conditions physiologiques. Par exemple, le dipeptide peut être lié à un site actif d'un médicament pour former un promédicament et/ou le dipeptide peut comporter un polymère de retard pour séquestrer une composition injectable comportant le complexe au point d'administration.
PCT/US2011/039755 2010-06-24 2011-06-09 Agents médicinaux liés à un dipeptide WO2011162968A1 (fr)

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US13/697,050 US20130137849A1 (en) 2010-06-24 2011-06-09 Dipeptide linked medicinal agents
CA2796879A CA2796879A1 (fr) 2010-06-24 2011-06-09 Agents medicinaux lies a un dipeptide
JP2013516595A JP6101202B2 (ja) 2010-06-24 2011-06-09 ジペプチド結合医薬剤
RU2013102991/04A RU2013102991A (ru) 2010-06-24 2011-06-09 Связанные с дипептидом лекарственные средства
MX2012014573A MX2012014573A (es) 2010-06-24 2011-06-09 Agentes medicinales unidos a dipeptido.
CN201180031396.0A CN102958533B (zh) 2010-06-24 2011-06-09 二肽连接的药剂
KR1020127033201A KR20130083843A (ko) 2010-06-24 2011-06-09 디펩티드 연결된 약제
EP11798607.5A EP2588127A4 (fr) 2010-06-24 2011-06-09 Agents médicinaux liés à un dipeptide
US15/016,959 US20160158375A1 (en) 2010-06-24 2016-02-05 Dipeptide linked medicinal agents

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US9845287B2 (en) 2012-11-01 2017-12-19 Aileron Therapeutics, Inc. Disubstituted amino acids and methods of preparation and use thereof
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US10213477B2 (en) 2012-02-15 2019-02-26 Aileron Therapeutics, Inc. Peptidomimetic macrocycles
US10227380B2 (en) 2012-02-15 2019-03-12 Aileron Therapeutics, Inc. Triazole-crosslinked and thioether-crosslinked peptidomimetic macrocycles
US10253067B2 (en) 2015-03-20 2019-04-09 Aileron Therapeutics, Inc. Peptidomimetic macrocycles and uses thereof
US10301351B2 (en) 2007-03-28 2019-05-28 President And Fellows Of Harvard College Stitched polypeptides
US10471120B2 (en) 2014-09-24 2019-11-12 Aileron Therapeutics, Inc. Peptidomimetic macrocycles and uses thereof
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CN112996486A (zh) 2017-07-17 2021-06-18 K·罗伊兹曼 包含细胞穿透肽的治疗剂的局部递送用于治疗年龄相关性黄斑变性及其它眼病

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US10301351B2 (en) 2007-03-28 2019-05-28 President And Fellows Of Harvard College Stitched polypeptides
US9957299B2 (en) 2010-08-13 2018-05-01 Aileron Therapeutics, Inc. Peptidomimetic macrocycles
US9096684B2 (en) 2011-10-18 2015-08-04 Aileron Therapeutics, Inc. Peptidomimetic macrocycles
US9522947B2 (en) 2011-10-18 2016-12-20 Aileron Therapeutics, Inc. Peptidomimetic macrocycles
US10308699B2 (en) 2011-10-18 2019-06-04 Aileron Therapeutics, Inc. Peptidomimetic macrocycles
US10213477B2 (en) 2012-02-15 2019-02-26 Aileron Therapeutics, Inc. Peptidomimetic macrocycles
US10227380B2 (en) 2012-02-15 2019-03-12 Aileron Therapeutics, Inc. Triazole-crosslinked and thioether-crosslinked peptidomimetic macrocycles
US9845287B2 (en) 2012-11-01 2017-12-19 Aileron Therapeutics, Inc. Disubstituted amino acids and methods of preparation and use thereof
US10669230B2 (en) 2012-11-01 2020-06-02 Aileron Therapeutics, Inc. Disubstituted amino acids and methods of preparation and use thereof
US10471120B2 (en) 2014-09-24 2019-11-12 Aileron Therapeutics, Inc. Peptidomimetic macrocycles and uses thereof
US10253067B2 (en) 2015-03-20 2019-04-09 Aileron Therapeutics, Inc. Peptidomimetic macrocycles and uses thereof
US11779648B2 (en) 2020-07-22 2023-10-10 Novo Nordisk A/S Co-agonists at GLP-1 and GIP receptors suitable for oral delivery
WO2023139187A1 (fr) 2022-01-20 2023-07-27 Novo Nordisk A/S Promédicaments co-agonistes du récepteur glp-1/gip et leur utilisation
US11840560B2 (en) 2022-01-20 2023-12-12 Novo Nordisk A/S Prodrugs and uses thereof

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CN102958533B (zh) 2016-01-20
AR081984A1 (es) 2012-10-31
US20160158375A1 (en) 2016-06-09
TW201204392A (en) 2012-02-01
EP2588127A1 (fr) 2013-05-08
MX2012014573A (es) 2013-02-21
CN102958533A (zh) 2013-03-06
RU2013102991A (ru) 2014-08-10
JP6101202B2 (ja) 2017-03-22
US20130137849A1 (en) 2013-05-30
CA2796879A1 (fr) 2011-12-29
KR20130083843A (ko) 2013-07-23
JP2013531659A (ja) 2013-08-08
EP2588127A4 (fr) 2014-06-11

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